CN101356184A - Methods for assessing patients with acute myeloid leukemia - Google Patents

Abstract

Methods for treating cancer, and preferably hematological malignancy, patients include analyzing gene expression profiles and/or molecular markers of a patient to determine status and/or prognosis of the patient. The invention also provides methods of analyzing whether a non-relapsed or non-refractory patient is likely to respond to treatment with farnesyl transferase inhibitors (FTIs) and, optionally, other therapeutics. The methods are also useful for monitoring patient therapy and for selecting a course of therapy. Genes modulated in response to FTI treatment are provided and are used in formulating the profiles.

Description

The method that the patient who suffers from acute myelogenous leukemia is assessed

Incorporate appendix " sequence table " tabulation into this paper by reference in view of the above.

Background of invention

The present invention relates to diagnose, prejudge the method for (prognostics) and treatment acute myelogenous leukemia (AML) based on detection molecules mark and/or gene expression analysis.

Karyotyping at present is effectively providing aspect the prognostic value, although it also is used for AML hypotypes different on the characterization of biological.In addition, this sick pathogeny relates to such as the sudden change in the genes such as FLT3, c-KIT, AML1, GATA1, CEBPA and N-RAS.Clear and definite is can the patient with different risk of relapse to be divided into groups at the examination of FLT3 and CEBPA sudden change.Effective dangerous classification can make can adopt correct stem cell transplantation or other adjuvant therapy.There are two pieces of articles of publishing recently to describe the adult AML patient's of new diagnosis gene expression profile and the purposes in the prediction clinical effectiveness thereof.Bullinger et al. (2004); And Valk etal. (2004).These studies show that gene expression profile is how can further improve clinical outcome prediction.

Valk et al. (2004) assesses 285 patients (marrow or peripheral blood) on Affymetrix U133A chip.Patient's sample comprises the unusual and molecule abnormality of the cytogenetics of broad range.Only identify 16 clusters (cluster), show that AML may not have the heterogeneity of being thought before.The AML hypotype of determining on several clusters wherein and cytogenetics and the molecule conforms to well, therefore supports their purposes in the WHO categorizing system.Bullinger et al. (2004) and other small research of delivering have before also been observed these clusters.Schoch et al. (2002); Debernardi et al. (2003); And Kohlmann et al. (2003).Therefore these clusters needn't wonder that they are relevant with prognosis with known to prejudge caryogram related well.

Bullinger et al. (2004) employing cDNA array has been studied the express spectra (65 peripheral bloods and 54 marrow) from 116 adult patients.Except the work of Valk et al. (2004), they have also developed 133 gene classification and have been used to predict the dangerous clinical effectiveness of organizing of all cytogenetics.Use the training group of 59 duplicate samples and the test group of 57 duplicate samples, they confirm that this 133 gene is divided into bad the patient and good result group (p=0.006log rank; Odds ratio, 10,95%CI, 2.6-29.3).

It should be noted that genes identified in these two researchs only the predicted gene identified in the leukemia of children of part have overlapping.Yagi?et?al.(2003)。In addition, Bullinger et al. (2004) this group of prejudging gene of identifying and 3 predictions identifying recently gene that Zarnestra (tipifarnib) is replied does not have overlapping.U.S. Patent Application Serial Number 10/883,436.

Farnesyl transferase (FTase) mediation carbon farnesyl partly is covalently attached to C-terminal CAAX (C, halfcystine; A, aliphatic residue; X, any amino acid) the identification motif.Reiss?et?al.(1990)。This farnesylation is further processed by the isopentene group halfcystine of rupture 3 end amino acids and the C-terminal that methylates.The arrestin farnesylation has been eliminated the required correct Subcellular Localization of protein function.At first, carcinogenic Ras albumen is considered to the target of the antiproliferative effect of FTIs in carcinobiology.Reuter?et?al.(2000)。Yet, after this show all effects that can not explain Zarnestra to the inhibition of Ras farnesylation.For example, FTIs does not always require sudden change Ras albumen to exist to produce antitumor action.Karp?et?al.(2001)。In fact, although around the colony with high frequency ras sudden change (for example late period colorectum and carcinoma of the pancreas) design, comparing aspect response rate with placebo, early stage clinical study do not have significant difference.Van Cutsem et al. (2004); And Rao et al. (2004).

Several other albumen of farnesylation come into the picture into as candidate's target that can mediate the antitumorgienesis effect of FTIs, comprise little GTPase albumen Rho B, centromere protein CENP-E and CENP-F, Protein-tyrosine-phosphatase PTP-CAAX and structural nuclear lamina protein A of nuclear membrane and B.Suppress these proteic farnesylations and may cause the antiproliferative effect of FTIs and adjust several important signaling molecules indirectly, comprise TGFbRII, MAPK/ERK, PI3K/AKT2, Fas (CD95), NF and VEGF.Adnane et al. (2000); Morgan et al. (2001); Jiang et al. (2000); Na et al. (2004); Takada et al. (2004); And Zhang et al. (2002).Regulate the adjusting that these signal pathways cause cell growth, propagation and apoptosis.Thereby FTIs may have complicated restraining effect to several cell incidents and approach.

The current method that does not have to determine these patients' state or predict total survival rate.

Summary of the invention

The invention provides a kind of method, wherein use one or more genetic markers to predict to suffer from acute myelogenous leukemia the patient's of (AML) the method for prognosis (prognosis).These marks can use or unite use separately, and this depends on the type of pharmacological agent.

The invention provides method by assessment acute myelogenous leukemia (AML) state, this is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, wherein above or the expression level that is lower than the marker gene of predetermined threshold level indicate the AML state.

The invention provides method by stages to acute myelogenous leukemia (AML) patient, this is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, wherein above or the expression level that is lower than the marker gene of predetermined threshold level indicate the AML survival rate.

The invention provides the method for definite acute myelogenous leukemia (AML) patient treatment scheme, this is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, the expression level that wherein surpasses or be lower than the marker gene of predetermined threshold level is enough to indicate to the replying of therapy, so that the doctor can determine to recommend to be used to provide the degree and the type of the therapy of appropriate therapeutic.

The invention provides treatment acute myelogenous leukemia (AML) patient's method, this is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, the expression level that wherein surpasses or be lower than the marker gene of predetermined threshold level indicates replying therapy, if and they have respondent spectrum, then treat described patient with adjuvant therapy.

The invention provides the method for the dangerous height of definite acute myelogenous leukemia (AML) death, this is undertaken by following process: obtain biological sample from AML patient, and measure and carry out corresponding to being selected from the relevant biomarker of those marker gene in the table 3, the expression level that wherein surpasses or be lower than the marker gene of predetermined threshold level indicates dead danger, so that the doctor can determine the degree and the type of the therapy of being recommended.

The invention provides and generate acute myelogenous leukemia (AML) and prejudge patient's method of reporting, it is by determining in the aforesaid method any result, and prepares to show that the report of this result and the report of generation thus finishes.

The invention provides and in biological sample, measure to determine the test kit of acute myelogenous leukemia (AML) prognosis, comprise the isolated nucleic acid sequences that is used to detect one group of gene, their complementary sequence or the material of its part, this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

The invention provides the goods of assessment acute myelogenous leukemia (AML) state, it comprises: be used to detect the isolated nucleic acid sequences of one group of gene, their complementary sequence or the material of its part, this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

The invention provides microarray or gene chip and be used to carry out aforesaid method.

The invention provides diagnosis/prognosis bag (portfolio), it comprises the isolated nucleic acid sequences of one group of gene, their complementary sequence or its part, and this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

Brief Description Of Drawings

Unsupervised (unsupervised) cluster of the AML patient of Fig. 1 recurrence and refractory.This tree derivation shows 58 recurrences or refractory AML patient's unsupervised k-mean cluster, and wherein each row is represented the patient, and each row is represented gene.By expression of gene level among the patient is calculated each expression of gene ratio divided by all other patients' mean value.Colour band shows variation multiple (log ₂).Redness raises, and blueness is reduced.White shows no change.Show and have 6 main clusters.

The real-time RT-PCR of Fig. 2 .2 gene.Measure AHR and AKAP13 by real-time RT-PCR.HPRT or PBGD crt gene are used for stdn genetic expression value.Error line is a standard deviation.The microarray data mapping that the value that obtains is corresponding relatively, the line linearity regression analysis of going forward side by side.

Fig. 3 shows the predictor of AKAP13 gene.Illustration (panel) A shows that the 2x2 that obtains from the LOOCV that adopts AKAP13 to express to carry out respondent (R) and non-responder (NR) as sorter (classifier) shows.Illustration B shows 58 same patients AKAP13 expression values.The P value shows genetic expression, and there were significant differences between the mean value of respectively replying group.Illustration C shows the Kaplan-Meier curve that generates from the patient who is divided into respondent and non-responder by the AKAP13 gene.

Fig. 4 provides the evaluation to the smallest group of prediction indication thing.In illustration A, adopt 100% susceptibility to carry out LOOCV.Tested the independently sorter that contains 1 to 19 kind of gene.Error rate mapping to the result.Illustration B shows that the 2x2 that obtains from the LOOCV that adopts 3 genetic markers to carry out respondent (R) and non-responder (NR) as the classification implements shows.Illustration C shows the score that generates from this 3 gene sorter.The P value shows genetic expression, and there were significant differences between the group respectively replying.Illustration D shows the Kaplan-Meier curve that generates from the patient who is divided into respondent and non-responder by this 3 genetic marker.Also show the intermediate value survival time.

Fig. 5. the respondent that is divided into prediction by 3 genetic markers and non-responder's patient is carried out Kaplan-Meier analyze.Shown and be defined as the non-responder clinically but adopt this 3 genetic marker to be categorized as respondent's patient's survivorship curve.Also pointed out the intermediate value survival time.

Fig. 6 has described AKAP13 crossing in AML clone and has expressed.Cell counting is normalized to the culture (showing at-12 log unit places) that does not have medicine, to provide the per-cent with respect to contrast.Error line is pointed out the average mistake.Hollow data points shows the result that the second time, experiment obtained by research higher concentration medicine.

Fig. 7 provides the model of FTI effect among recurrence or the refractory AML.A. in the respondent, IL3RA and AKAP13 gene are low expresses, and ras, RhoA and lamin B approach are reduced.The rise of RhoH has been caused strengthening the inhibition of pair cell path for transformation.Altogether, this makes that the antitumor generation of FTI can be more effective.B. opposite express spectra seems to make compensation approach to be expressed among the non-responder.

Fig. 8 .Zarnestra predicted gene mark is more useful than independently prejudging genetic marker.In illustration A, the row representative is from the AML sample that recurs or the refractory patient obtains, and row is represented 103 167 probe groups in 133 prognosis genes identifying corresponding to people such as Bullinger, sorts according to hierarchical clustering.Illustration B shows the Kaplan-Meier survival rate evaluation that the patient by the cluster definition is organized.In illustration C, 3 gene sorters have been used for prejudging group Bullinger tag definitions good and bad and have identified the Zarnestra respondent.Shown the Kaplan-Meier survivorship curve that in good (Zn+. cluster 1) and bad (Zn+. cluster 2) prognosis group, is accredited as the Zarnestra respondent.Pointed out the intermediate value survival time of each group.

Fig. 9 is a schema, shows how gene from people such as Bullinger (2004) mates 167 probe groups (103 unique genes) on the Affymetrix U133A chip.

Figure 10 shows that 167 probe groups are marked at the application among recurrence or the refractory AML patient.In illustration A, the row representative is from the AML sample that recurs or the refractory patient obtains, and row is represented 103 167 probe groups in 133 prognosis genes identifying corresponding to people such as Bullinger (2004), sorts according to hierarchical clustering.Illustration B shows the Kaplan-Meier survival rate evaluation that the patient by the cluster definition is organized.

Figure 11 provides the comparison of prejudging with Zarnestra predicted gene mark.Illustration A shows the good and bad Kaplan-Meier survivorship curve of prejudging cluster by the subset definition of 103 Bullinger et al. (2004) gene.Illustration B shows by the good and bad KBplBn-Meier survivorship curve of prejudging cluster of predicting the 3 genetic markers definition that Zarnestra is replied.Illustration C shows by the good and bad KBplBn-Meier survivorship curve of prejudging cluster among the further stratified illustration A of prediction 3 genetic markers.Illustration D shows that prediction has poor prognosis and do not reply the Kaplan-Meier survivorship curve of the patient of Zarnestra to all the other patients.

Figure 12 identifies the group of the minimum of prediction indication thing.A). carry out LOOCV, select susceptibility 100%, specificity 40% and multiple to change＞2 gene.Tested the independent sorter that contains by 1 to 8 gene of AUC grading.Drawn result's error rate.B) 2x2 that obtains from the LOOCV that adopts AKAP13 to carry out respondent (R) and non-responder (NR) as sorter shows.C) the genetic expression value of AKAP13.The P value shows genetic expression, and there were significant differences between the group replying.D) the Kaplan-Meier curve that generates from the patient who is divided into respondent and non-responder by AKAP13.Also shown the intermediate value survival time.

Figure 13 shows the sketch plan of gene expression analysis.

Figure 14 shows: after the Zarnestra treatment stopped, the AML sample was kept total changes in gene expression of FTI mediation.

Figure 15 shown in new diagnosis AML the prediction express spectra and to the test of prediction sorter.

Figure 16 has shown that 6 gene sorters are classified to new diagnosis AML.

Detailed description of the invention

Being described in the past the subgroup that has the gene of prejudging value among the AML of new diagnosis shows in the recurrence of targeted molecular therapy (Zarnestra) treatment and intractable AML patient useful here. The current method of replying that is used for predicting to farnesyl transferase inhibitor (such as Zarnestra). In addition, the existing method that is used for understanding AML patient's prognosis only limits to histological subtypes and caryogram, but neither is the desirable mark of determining clinical effectiveness. Current label has been expanded conventional art by providing to the classification of high-risk and low dangerous patient.

Sequence number is that 10/883,436 U.S. Patent application proves that the AML patient of 3 gene classifiers (comprising AHR, AKAP13 and MINA53) prediction recurrence, refractory has extremely low error rate to replying of Zarnestra. Also can see this point when staying five methods (leave-five-out) cross validation. When adding more polygenes, error rate increases, and shows that episome introduced noise to this classification. For 3 gene separating methods, LOOCV is presented at has 86% sensitiveness and 70% specificity when overall diagnosis accuracy is 74%. Kaplan-Meier analyzes and shows that also there is significant difference in survival rate between respondent's group of predicting and non-responder's group. In addition, when the non-responder of the non-responder of misclassification and correct classification was compared, the non-responder of misclassification had shown preferably overall survival.

Be the competitive farnesyl transferase inhibitor (FTI) of non-plan peptide that can be oral, it has demonstrated the propagation that suppresses the various human tumor cell line in vitro and in vivo. End et al. (2001); With Cox et al. (2002). The I clinical trial phase of Zarnestra has shown that the patient to suffering from refractory or recurrence acute myelogenous leukemia has 32% response rate. Karp et al. (2001). Also observe the activity to myelodysplastic syndrome (MDS) (Kurarock et al.2004), Huppert's disease (MM) (Alsina et al. (2003)) and chronic myelogenous leukemia (CML) in the clinical research in early days. Cortes et al. (2003). Alleviate the bone marrow damage be defined as less than 5% fully, neutrophil count greater than 1000/ μ L, platelet count is less than 100,000/ μ L and do not have the outer disease of marrow. Work by the CKIs farnesylation although know FTIs, still do not know the antitumaous effect implication aspect the green blood malignant tumour of which gene and Zarnestra. Microarray technology be so that can measure simultaneously the steady-state mRNA level of thousands of genes, thereby represented for the identification of the gene relevant with the FTI effect and the effective tool of gene approach. Therefore, whole gene expression monitoring is used in the AML of recurrence and refractory the 2 phases clinical research of Zarnestra is predicted in haematological malignancies this FTI is had the gene of replying to identify.

Only in genome, exist might expressing protein or the nucleotide sequence of peptide can not determine whether albumen or peptide express in given cell. Given can expressing protein or whether the gene of peptide or transcribe rna is done like this and such expression or transcribe the degree (if any) of generation, determined by the Various Complex factor. Yet measuring gene expression can provide about the useful information of replying of cell to given stimulation, for example to introducing replying of medicine or other therapeutic agent. The relative indication of gene activation or level of deactivation can be found in such gene expression profile. In some cases, the existence of molecular marker also can be independently or be provided useful information about treatment validity by means of gene expression information. Gene expression profile of the present invention and molecular marker are used for identifying and treatment AML patient.

The cancer that comprises haematological malignancies results from the sudden change of several genes usually. The cancer of same type may originate from one or more sudden changes or with corresponding to one or more sudden changes, but these sudden changes may be different from the sudden change of another patient with same type cancer. In fact, different kinds of molecules basis is usually arranged under identical cancer, this affects a patient with some therapies of observing but another patient that may not similarly affect the cancer of suffering from same type conforms to. In addition, from the viewpoint of diagnosis, the existence of specific sudden change (such as transposition, deletion or SNPs) can have remarkable meaning. In some cases, such molecular marker they oneself be exactly diagnosis, prognosis or determine the useful indicator that treatment is replied. Can be with especially true for the related situation of the reacting phase of particular treatment for molecular mutation.

Biomarker is any indicant of the expression of the marker gene pointed out. This indication can be directly or indirectly, and compares with interior mark, normal structure or another cancer, weighs the enhancing of the expression of gene under the given physiological parameter or weakens. Biomarker includes but not limited to nucleic acid (express to strengthen and weaken and directly and indirectly). Use nucleic acid can comprise any method well known in the prior art as biomarker, include but not limited to weigh the low or hyper-methylation of DNA amplification, RNA, little RNA, loss of heterozygosity (LOH), SNP (SNPs, Brookes (1999)), microsatellite DNA, DNA. Use albumen can comprise any method well known in the prior art as biomarker, include but not limited to weigh quantity, activity is modified such as glycosylation, phosphorylation, ADP-ribosylation, ubiquitin etc. SABC (IHC). Other biomarker comprises imaging, cell count and apoptosis mark.

Given gene provided herein is those relevant with specific tumors or types of organization. Marker gene may with many type of cancer and relevant; but as long as adopt special algorithm for lung carcinoma cell described herein to show that the expression of this gene is relevant fully with a kind of tumour or types of organization to be identified; then this gene can be used among claimed the present invention, to determine cancerous state and prognosis. Many genes of known and one or more related to cancer in the prior art. The invention provides preferred marker gene and even preferred marker gene combination. These have a detailed description in this article.

When it contained this sequence, marker gene was corresponding to the sequence by SEQ ID appointment. Gene section or fragment be corresponding to the sequence of this gene, and condition is that the part of its described sequence that contains or its complementary series are enough to make it to be differentiated sequence for this gene. Gene expression product is corresponding to this sequence, and condition is its RNA, mRNA or cDNA and the composition that contains this sequence (for example probe) hybridization, or with regard to peptide or albumen, it is by this mRNA coding. Gene expression product section or fragment be corresponding to the sequence of this gene or gene expression product, and condition is that the part of its described gene expression product that contains or its complementary series are enough to make it to be differentiated sequence for this gene or gene expression product.

This specification is described and the inventive method, composition, goods and the kit of opinion comprise one or more marker genes. " mark " or " marker gene " that spread all over the use of this specification refers to following gene and gene expression product, and it is corresponding to excessively expressing and owing to express any gene relevant with tumour or types of organization. Preferred marker gene is described in more detail in the table 8.

The invention provides the method for assessment acute myelogenous leukemia (AML) state, it is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to being selected from the relevant biomarker of those marker gene in table 3, table 4, table 5, table 7, table 8 or the table 9, wherein above or the expression that is lower than the marker gene of predetermined threshold level indicate the AML state.

The invention provides the method by stages to acute myelogenous leukemia (AML) patient, it is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to being selected from the relevant biomarker of those marker gene in table 3, table 4, table 5, table 7, table 8 or the table 9, wherein above or the expression that is lower than the marker gene of predetermined threshold level indicate the AML survival rate.

The invention provides the method for definite acute myelogenous leukemia (AML) patient treatment scheme, it is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, the expression level that wherein surpasses or be lower than the marker gene of predetermined threshold level is enough to indicate to the replying of therapy, so that the doctor can determine to recommend to be used to provide the degree and the type of the therapy of appropriate therapeutic.

The invention provides treatment acute myelogenous leukemia (AML) patient's method, it is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, the expression level that wherein surpasses or be lower than the marker gene of predetermined threshold level indicates replying therapy, if and they have respondent spectrum, then treat described patient with adjuvant therapy.

The invention provides the method for the dangerous height of definite acute myelogenous leukemia (AML) death, it is undertaken by following process: obtain biological sample from AML patient, and measure with corresponding to those the relevant biomarker of marker gene that is selected from the table 3, the expression level that wherein surpasses or be lower than the marker gene of predetermined threshold level is enough to indicate dead danger, so that the doctor can determine the degree and the type of the therapy of being recommended.

Method provided herein also may comprise, contains or utilize the measurement of at least a expression of gene level that composing type ground in sample is expressed.Preferably, method provided herein produces the specificity at least about 40%.Preferably, method provided herein produces the susceptibility at least about 80%.Preferably, method provided herein produces the p value less than 0.05.

Method provided herein can be undertaken by measure genetic expression on microarray or gene chip.This microarray can be cDNA array or oligonucleotide array, and may also contain mark reagent in one or more.

Method provided herein can be undertaken by measuring genetic expression, and this genetic expression is realized by the nucleic acid amplification to the RNA that extracts from sample that is undertaken by polymerase chain reaction (PCR).This PCR can be reverse transcriptase polymerase chain reaction (RT-PCR), and can contain mark reagent in one or more.

Method provided herein can be undertaken by the albumen of measuring or detect by this genes encoding.This albumen can detect with being specific to this proteic antibody.

Method provided herein can be undertaken by the feature of this gene.Feature includes but not limited to DNA cloning, methylates, sudden change and allelic variation.

The invention provides generation acute myelogenous leukemia (AML) prognosis patient method of reporting, it is finished by any the result and the report of preparation this result of demonstration and the report of generation thus in definite aforesaid method.This report may contain to patient result and/or with respect to patient group's dangerous probability and/or possibility or to the assessment of the reaction of chemotherapy.

The invention provides and in biological sample, measure to determine the test kit of acute myelogenous leukemia (AML) prognosis, it comprises: be used to detect the isolated nucleic acid sequences of one group of gene, their complementary sequence or the material of its part, this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.This test kit also can contain and is useful on reagent and/or the medium that carries out microarray analysis, measures described nucleotide sequence, their complementary sequence or its part by them.

The invention provides the goods of assessment acute myelogenous leukemia (AML) state, it comprises: be used to detect the isolated nucleic acid sequences of one group of gene, their complementary sequence or the material of its part, this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.These goods also can contain and are useful on reagent and/or the medium that carries out microarray analysis, measure described nucleotide sequence, their complementary sequence or its part by them.

The invention provides the microarray or the gene chip that are used to carry out aforesaid method.This microarray may contain the isolated nucleic acid sequences of one group of gene, their complementary sequence or its part, and this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.Preferably, this microarray provides measurement or the sign that at least 1.5 times of mistakes are expressed or owed to express.Preferably, this microarray provides the measurement with statistically evident p value to crossing to express or to owe to express.More preferably, this p value is less than 0.05.This microarray can be any known microarray in the prior art, includes but not limited to cDNA array or oligonucleotide array, and mark reagent in can also containing.

The invention provides diagnosis/prognosis bag, it comprises the isolated nucleic acid sequences of one group of gene, their complementary sequence or its part, and this group gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.Preferably, this measurement or be characterized by at least 1.5 times of mistakes and express or owe and express.Preferably, this measurement has statistically evident p value to crossing to express or owe to express to provide.More preferably, this p value is less than 0.05.

The preferred method of setting up gene expression profile comprises the amount of the RNA that determines that the gene by codified albumen or peptide produces.This finishes by reverse transcriptase PCR (RT-PCR), competitive RT-PCR, real-time RT-PCR, difference demonstration RT-PCR, Northern engram analysis and other dependence test.Although might adopt various PCR reaction to carry out these technology, the complementary DNA (cDNA) that produced by mRNA or complementary RNA (complementary RNA) and it is analyzed by microarray preferably increase.Many different array structures and their production method are known to those skilled in the art, and description is arranged in United States Patent (USP), for example: 5,445,934; 5,532,128; 5,556,752; 5,242,974; 5,384,261; 5,405,783; 5,412,087; 5,424,186; 5,429,807; 5,436,327; 5,472,672; 5,527,681; 5,529,756; 5,545,531; 5,554,501; 5,561,071; 5,571,639; 5,593,839; 5,599,695; 5,624,711; 5,658,734; And 5,700,637.

Microarray technology makes the steady-state mRNA level can measure thousands of genes simultaneously, thus for identify to uncontrolled cell proliferation such as starting, stagnate or effect such as adjusting providing effective instrument.Two kinds of current being used widely of microarray technology.First kind is the cDNA array, and second kind is the oligonucleotide array.Though there are differences aspect these chips of structure, all basically downstream data analyses all are identical with output.The result of these analyses is generally the measurement of the strength of signal that the probe from mark is received, and the probe of this mark is used for the sequence from sample detection cDNA, and this cDNA sequence hybridizes on this microarray in known location.Usually, the intensity of this signal and the amount of cDNA are proportional, thus also with sample cell in the amount of the mRNA that expresses proportional.A lot of such technology are obtainable and useful.The preferred method of determining genetic expression can be in United States Patent (USP) 6,271,002; 6,218,122; 6,218,114; And find in 6,004,755.

Carry out the expression level analysis by more such strength of signal.This preferably finishes at the neutralize rate matrix of its expression intensity in control sample of test sample by generating gene.For example, will compare from the expression intensity that the genetic expression intensity of illing tissue and optimum or healthy tissues from same type generate.The ratiometer Benq of these expression intensities is because of being expressed in the variation multiple between test and the control sample.

The also available multiple mode of gene expression profile shows.Modal method is that primary fluorescence intensity or rate matrix are arranged in the tree derivation (dendogram), and wherein test sample, line display gene are shown in tabulation.Carry out data ordering by described mode hereinafter, described mode makes that the gene with similar express spectra is closer to each other.Each expression of gene ratio manifests as color.For example, ratio may manifest at the spectrographic blue portion less than 1 (downward modulation), and ratio may manifest in the red part of spectrographic greater than 1 (showing rise).Can buy and be used for showing these data computing machine software, Silicon Genetics for example, " GENESPRING " of Inc., Partek, " DISCOVERY " of Inc and " INFER ".

Measuring under the situation of protein level with mensuration genetic expression, as long as it produces enough specificity and susceptibility, any currently known methods of the prior art all is fit to.For example, can measure protein level by being attached to the amount that is specific to this proteic antibody or antibody fragment and measures antibody binding proteins.Antibody can be detected with promotion by radioactive, fluorescence or other detectable reagent mark.Detection method includes but not limited to enzyme-linked immunosorbent assay (ELISA) and immunoblot assay.

(modulated) mark adjusted that is used for the inventive method is described in embodiment.Differentially expressed gene raises in having the patient that various lung cancer prejudge or downward modulation.Last mediation downward modulation is relative terms, refers to find that gene expression amount has detectable difference (surpassing the influence of noise of the system that is used to measure it) with respect to certain baseline values.In this case, baseline values is determined based on algorithm.Adopt identical measuring method so, the goal gene in the diseased cells raises or downward modulation with respect to baseline values.

The mensuration of the genetic expression state of pair cell also can be determined normal/abnormal tissue distribution, is used for adopting the diagnosis such as the technology of immunohistochemical analysis (IHC).Any known method can both be used in the prior art, for example with regard to LBC oncogene, can unite use with prepare for IHC research FF and through formalin fixed, paraffin-embedded tissue block at the proteic antibody of LBC.Each tissue block can be made of residual " pulverizing " tumour of 50mg.

In brief, freezing microtome section can prepare like this: under the room temperature, and in the phosphate buffered saline (PBS) in little plastic capsule (PBS), the freezing and pulverizing tumour of rehydration 50ng; By the centrifugation particle; They are resuspended in the embedding medium (OCT) of viscosity; The counter-rotating capsule is also once more by centrifugation; Rapid condensation in-70 ℃ iso-pentane; The cutting plastic capsule also takes out the cylindric tissue of refrigerated; To organize right cylinder to be fixed on the cryogenic thermostat slicing machine chuck; And cutting-out contains the 25-50 serial section of intact tumor cells.Permanent section can be by similar method preparation, and it relates to the sample of rehydration 50mg in the plastics micro-centrifuge tube; Precipitation; Resuspension was used for fixing in 4 hours in 10% formalin; Washing/precipitation; Resuspension in 2.5% agar; Precipitation; Cooling in frozen water is so that the agar sclerosis; From pipe, take out and organize agar block; Filter and this piece embedding is advanced in the paraffin, and the continuous permanent section of downcutting as many as 50.

For IHC measured, described section was coated with lock solution, and it contains: 3% bovine serum albumin (BSA) or other closed reagent among the PBS.Described closed reagent comprises non-specific serum or milk powder.Allow sealing at room temperature carry out 1 hour.With containing 3%BSA, 0.1%TritonX ^TM-100 and the PBS of uncle's octylphenoxy polyethoxyethanol with the proteic antibody of the anti-LBC of 1: 100 dilution proportion.The sample section covers 16hr for 4 ℃ with antibody-solutions usually.Time length and temperature condition can change according to selected antibody and the material of being tested.Optimal conditions is rule of thumb determined.Section through antibody treatment is washed three times in PBS subsequently, and each 15 minutes, to remove unconjugated antibody, the second antibody with PBS that contains 3%BSA and dilution in 1: 2000 covered then.This second antibody can be coupled to the enzyme that adds lustre to, as: horseradish peroxidase, alkaline phosphatase, fluorescein isothiocyanate or other enzyme that is fit to.Perhaps, this second antibody can be coupled to vitamin H, and unites use with the avidin of chromophoric group mark.

The method that another exemplary detection gene exists is for by in situ hybridization.Usually, in situ hybridization comprises following key step: (1) fixes tissue or biological structure to be analyzed; (2) prehybridization is handled this biological structure, with the accessibility (accessibility) of increase target DNA, and reduces non-specific binding; (3) make nucleic acid mixture and biological structure or the tissue in nucleic acid hybridization; (4) post-hybridization washing is to remove unconjugated nucleic acid fragment in the hybridization; And (5) detect the nucleic acid fragment of hybridization.The reagent and the working conditions that are used for these each steps of step change with application-specific.

In this case, comprise at least a can with the hybridization solution of the detected nucleic acid probe of gene (at its chromosomal foci) hybridization under hybridization conditions with cells contacting.Detect any hybridization then, and compare with the predetermined crossing pattern that comes from normal or control cells.Preferably, this probe is α-centromeric probe.Such probe can buy from a lot of sources (for example, from VisysInc., Downers Grove, IL).In preferred embodiments, this hybridization solution contains multiple probe, and they are specific to the zone on the karyomit(e), and these zones are corresponding to constituting chimeric transposition (for example, 15q24-25).

The crossing scheme that is applicable to the inventive method for example is expressed in Albertson (1984); Pinkel (1988); EP No.430,402; And Methods in Molecular Biology, Vol.33:InSitu Hybridization Protocols, Choo, ed., Humana Press, Totowa is among the NJ (1994) etc.In a kind of especially preferred embodiment, use the crossing scheme of Pinkel et al. (1998) or Kallioniemi (1992).The method of optimizing hybridization conditions be known (referring to, for example, Tijssen (1993) Laboratory Techniques in Biochemistry andMolecular Biology, Vol.24:Hybridization With Nucleic Acid Probes, Elsevier, NY).

In a kind of preferred embodiment, (for example, C-TAB) or closed reagent (for example, sperm DNA, cot-1DNA or the like), reduce non-specific binding and reduce background signal by during hybridizing, using scale remover.Preferably, hybridization (for example, cot-1DNA) is carried out under the existence at about 0.1 to 0.5mg/mL DNA.

Described probe can be by any method preparation well known in the prior art, comprises synthetic or cultivates in the biology host.Synthetic method includes but not limited to that oligonucleotide is synthetic, riboprobes and PCR.

Can come the described probe of mark with detectable mark by any method well known in the prior art.The method of label probe comprises random start, end mark, PCR and nick translation.Carry out under enzymatic the 4th kind of Nucleotide that is marked at nucleic acid polymerase, three kinds and does not have the Nucleotide of mark and mark, wherein the 4th kind of Nucleotide is direct mark, contains the connecting arm that is useful on the additional marking thing that perhaps the 4th kind of Nucleotide is attached to the combinative haptens of the binding molecule that is labeled or other molecule.The direct marker that is fit to comprises radioactively labelled substance, and as 32P, 3H and 35S, and the non-radioactive marker, fluorescent marker for example is as fluorescein, Texas is red, AMCA is blue, lucifer is yellow, rhodamine or the like; With the detectable cyanine dyes of visible light; Enzyme, or the like.Marker also can be by hydrosulphite mediation transamination from chemically adding to advance dna probe or directly adding to come between synthesis phase at oligonucleotide.

Fluorescent marker can easily be attached to the Nucleotide with activation connecting arm that adds to advance probe.Probe can be labeled indirectly by above disclosed method, it is by introducing covalently bound Nucleotide to haptens or other molecule such as vitamin H or digoxin (digoxygenin), and carry out sandwich hybridization with the antibody that is labeled at this haptens or other molecule, or, use the avidin that is coupled to detectable with regard to vitamin H.Antibody and avidin can be with fluorescent markers or with enzyme labelling thing such as alkaline phosphatase or horseradish peroxidase coupling, so that they can be detected.Coupling avidin and antibody can be from (Burlingame, CA) (Indianapolis, company IN) buys with Boehringer Mannheim such as VectorLaboratories.

By the substrate of enzyme is provided, described enzyme can detect by colorimetric reaction.In the presence of various substrates, produce distinct colors by reaction, these colors can be shown to detect multiple probe respectively.Any substrate well known in the prior art can use.The substrate that preferably is used for alkaline phosphatase comprises 5-bromo-4-chloro-3-indoles phosphoric acid and nitroblue tetrazolium (NBT) (NBT).The substrate that preferably is used for horseradish peroxidase is diaminobenzophenone (DAB).

It is long to be applicable to that the fluorescently-labeled probe of in-situ hybridization method of the present invention is preferably 150 to 500 Nucleotide.Probe can be DNA or RNA, preferred DNA.

The hybridization of detectable probe and cell is carried out under concentration and probe concentration 0.1-500ng/ μ L, preferred 5-250ng/ μ L.Hybridization mixture preferably contains the denaturing agent such as methane amide.Usually, hybridization is carried out preferred 32 ℃-40 ℃, most preferably 37 ℃-38 ℃ under 25 ℃-45 ℃.Hybridizing the required time is about 0.25-96 hour, more preferably 1-72 hour, and most preferably 4-24 hour.Hybridization time is based on concentration and probe concentration and hybridization solution content and change, and this solution may contain accelerator,, trialkyl ammonium salts conjugated protein as hnRNP, lactan etc.Subsequently with containing the solution washing slide glass of denaturing agent such as methane amide and lower concentration chlorination sodium, perhaps in any solution of removing not combination and mismatch probe, wash.

The concentration of temperature and salt changes with required hybridization stringency.For example, the washing of high stringency can be carried out at 42 ℃-68 ℃, and the scope that middle stringency can 37 ℃-55 ℃ is carried out, and low stringency can carry out at 30 ℃-37 ℃.The salt concn that is used for high stringency washing can be 0.5-1 SSC (0.15M NaCl, 0.015M Trisodium Citrate) doubly, and medium stringency can be 1-4 times, and low stringency can be 2-6 SSC doubly.

If desired, detect incubation step and preferably in damp camera, carry out, more preferably, most preferably under 37-38 ℃, carry out at 25 ℃-38 ℃ at 23 ℃-42 ℃.The reagent of mark preferably dilutes in the solution that contains such as the closed reagent of BSA, skim-milk etc.Dilution may be at 1: 10 to 1: 10, changes more preferably 1 between 000: 50-1: 5,000, and most preferably 1: 100-1: 1,000.Slide glass or other solid support should be washed between each incubation step to remove excessive reagent.

Subsequently,, slide glass is fixed and used microscopical analysis, or, analyze by being exposed to autoradiographic film for radioactively labelled substance for the visible detectable.With regard to fluorescent marker, preferably slide glass is immersed and contain in the solution of anti-cancellation reagent, and adopt the fluorescent microscope analysis.Can check that a plurality of nuclears are to increase accuracy in detection.

In addition, the mensuration to LBC oncogene expression of results can also be used to determining whether LBC oncogene sudden change has taken place.Most preferably, like this be determined as immunoassay.On their the simplest direct meaning, immunoassay is in conjunction with measuring.Some preferred immunoassaies are various types of enzyme-linked immunosorbent assay well known in the prior art (ELISAs) and radioimmunoassay (RIA).Adopt the IHC detection of tissue slice also to be particularly useful, in situ hybridization and enzyme immunoassay also are like this.

In a kind of exemplary enzyme-linked immunosorbent assay, protein specific antibody is fixed to the selected surface that presents the albumen avidity, for example polystyrene microtiter plates hole.Then, will contain required antigenic test composition such as clinical sample and be added to this hole.After combination and washing with the immunocomplex of removing non-specific combination, institute's bonded antigen can be detected.Usually realize detecting by adding the antibody that another kind is specific to required antigen and is connected to detectable.Such ELISA is simple " sandwich ELISA ".Also can be specific to required antigenic second antibody, add the 3rd antibody that second kind of antibody is had binding affinity and is connected to detectable subsequently and realize detecting by adding another kind.

The variation of elisa technique is known.In a kind of such variation, will contain required antigenic sample and be fixed to hole surface, contact with antibody of the present invention then.After combination and suitable washing, detect institute's bonded immunocomplex.When initial antigen-specific antibodies is connected to detectable, can directly detect immunocomplex.Equally, also can adopt the second antibody that first kind of antigen-specific antibodies had binding affinity and be connected to detectable to detect.

The detection of genetic expression be for the embodiment of determining AML prognosis or state in, it is most preferred using the genetic expression bag.The one group gene of gene bag for dividing into groups by following mode, described mode make the expressing information about them that is obtained provide the foundation for producing relevant clinically judgement (selecting as diagnosis, prognosis, treatment).In this case, the genetic expression bag is suitable for assisting AML patient is made the treatment decision.

In this article, the change of ill finger physical state, its interruption or interference maybe might be disturbed the intrinsic performance of body function, as uncontrolled cell proliferation takes place.When people's genotype or more phenotypic aspects were consistent with the existence of disease, it is diagnosed as suffered from disease.Yet, diagnose or the behavior of prognosis may comprise definite disease/state issues, as the possibility of determining recurrence, the type and the treatment monitoring of treatment.In treatment monitoring, express by icp gene in time, whether change or be not changed to the pattern more consistent to determine gene expression profile with healthy tissues, thus to given course of treatment effect make clinical judgment.

Gene can be grouped, and makes the judgement (selecting as diagnosis, prognosis, treatment) of being correlated with clinically for generation about complete expression of gene information in this group that is obtained that reliable basis is provided.This cover gene has constituted gene bag of the present invention.As most diagnostic marker, usually wish to use very a spot of marker just to be enough to make correct medical judgment.This has prevented to postpone owing to the analysis of products for further and the treatment that causes of duration of service and resource ineffectually.

A kind of method of setting up the genetic expression bag is by using optimal algorithm, as widely used average variance algorithm in setting up stock bag (stock portfolio).This method has a detailed description in the United States Patent (USP) of publication number 20030194734.In fact, present method requires to set up a cover input value (stock in financial application is to express with ionization meter) here, and it will optimize the return of value (for example signal of Sheng Chenging) that is used after reception, make the mutability minimum of return of value simultaneously.Many business software programs can be used for carrying out such operation.Preferably " WagnerAssociates Mean-Variance Optimization Application " is referred to as " Wagner software " in this manual.This software adopts determines efficiency frontier from the function of " Wagner AssociatesMean-Variance Optimization Library ", and the preferred package on the Markowitz meaning is a kind of selection.Use this type software requirement microarray data to be converted,, be used as the financial analysis purpose stock yield and the risk measurement that are used for itself when this software so that it can be taken as input value.

The process of selection gene bag can also comprise the application of heuristic rule.Preferably, such rule is based on biology with to the understanding of the technology that is used to generate clinical effectiveness and formulate.More preferably, they are applied to the output from optimization method.For example, adopt the gene bag of average variance method to select to can be applicable to suffer from the microarray data of numerous differentially expressed genes in the individuality of cancer.Described method is output as one group of gene of optimization, can be included in some genes of expressing in peripheral blood and the illing tissue.If the sample that is used to test obtains from peripheral blood, and for example differentially expressed some gene is also differentially expressed in peripheral blood in cancer, then can use heuristic (heuristic) rule, wherein the gene bag is selected from and has got rid of those efficiency frontiers differentially expressed in peripheral blood.Certainly, can by for example with this rule application during the data preliminary election, thereby before forming efficiency frontier, use this rule.

Can use unnecessary other heuristic rule relevant with the biology of being discussed.For example, can adopt such rule, promptly only the gene bag of specified percentage can be by specific gene or genome representative.The software that can buy as Wagner software, can easily adapt to such exploration.For example, when the factor except accuracy and accuracy (for example, expection licence fee) was influential to the hope that comprises one or more genes into, this was useful.

Gene expression profile of the present invention also can be united use with other the non-genetic diagnosis method that can be used for cancer diagnosis, prognosis or treatment monitoring.For example, in some cases, with the diagnosis capability of above-mentioned method based on genetic expression with will be useful from integrating such as the data of the conventional tag thing (for example tumour antigen 27.29 (" CA 27.29 ")) of serum protein marker.Have a series of such marker, comprise such analyte such as CA 27.29.In a kind of such method, regularly get blood from the patient of treatment, then above-mentioned serum markers a kind of carried out enzyme immunoassay.When the concentration of marker is represented the failure of tumor recurrence or treatment, obtain the sample source that is suitable for gene expression analysis.When suspicious lump exists, carry out fine needle aspiration, the gene expression of cells spectrum of this lump is taken from analysis as indicated above then.Perhaps, can be from the zone acquisition tissue sample contiguous with the tissue of removing tumour before.When other test generates ambiguously as a result the time, this method is particularly useful.

Test kit prepared in accordance with the present invention comprises the mensuration of the moulding that is used for definite gene expression profile.This can comprise measures required all or some material, as reagent and specification sheets and carry out the medium that biomarker is measured therein.

The method (comprising the method that is used to explain relevant biological pathway) of preferably setting up gene expression profile comprises the amount of the RNA that determines that the gene by codified albumen or peptide or transcribe rna produces.This preferably finishes by reverse transcription PCR (RT-PCR), competitive RT-PCR, real-time RT-PCR, difference demonstration RT-PCR, Northern engram analysis and other dependence test.Although might adopt various PCR reaction to carry out these methods, usually wish copy DNA (cDNA) that amplification is produced by mRNA or copy RNA (cRNA) and analyzed by microarray.Many different array structures and production method are known to those skilled in the art, and description is arranged in United States Patent (USP), for example: 5,445,934; 5,532,128; 5,556,752; 5,242,974; 5,384,261; 5,405,783; 5,412,087; 5,424,186; 5,429,807; 5,436,327; 5,472,672; 5,527,681; 5,529,756; 5,545,531; 5,554,501; 5,561,071; 5,571,639; 5,593,839; 5,599,695; 5,624,711; 5,658,734; And 5,700,637.

Microarray technology is measured the steady-state mRNA level of thousands of genes simultaneously, thereby is provided for identifying the effective tool of AML patient's gene expression profile.Two kinds of current being used widely of microarray technology.First kind is the cDNA array, and second kind is the oligonucleotide array.Though there are differences aspect these chips of structure, all basically downstream data analyses all are identical with output.The result of these analyses is generally the measurement of the strength of signal that the probe from mark is received, and the probe of this mark is used for the sequence from sample detection cDNA, and this cDNA sequence hybridizes on this microarray in known location.Usually, the amount of strength of signal and cDNA is proportional, thus also with sample cell in the amount of the mRNA that expresses proportional.A lot of such technology are obtainable and useful.Preferable methods can be in United States Patent (USP) 6,271,002; 6,218,122; 6,218,114; And find in 6,004,755.

Carry out the expression level analysis by more such intensity.This preferably finishes at the neutralize rate matrix of its expression intensity in control sample of test sample by generating gene.For example, will compare through the genetic expression intensity of the tissue of pharmacological agent and the expression intensity that homologue without pharmacological agent generates.The ratio of these expression intensities indicates the variation multiple of genetic expression between test and control sample.

Gene expression profile can show with multiple mode.Common method is that rate matrix is arranged in the tree derivation, the bright test sample of wherein tabulating, and row shows gene.Carry out data ordering in the following manner, described mode makes that the gene with similar express spectra is closer to each other.Each expression of gene ratio manifests as color.For example, ratio may manifest at the spectrographic blue portion less than 1 (showing downward modulation), and ratio may manifest in the red part of spectrographic greater than 1 (showing to raising).Can buy and be used for showing these data computing machine software, Silicon Genetics for example, " GENESPRING " of Inc., Partek, " DISCOVERY " of Inc and " INFER ".

Differentially expressed gene is in diseased cells or raise, and perhaps downward modulation is as knowing by inference by above-mentioned assessment genetic expression.Last mediation downward modulation is relative terms, refers to find that gene expression amount has detectable difference (surpassing the influence of noise of the system that is used to measure it) with respect to certain baseline values.In this case, baseline values is Normocellular measured genetic expression.Adopt identical measuring method so, the goal gene in the diseased cells raises or downward modulation with respect to baseline values.Preferably, be in harmonious proportion on the downward modulation level based on hybridization micro probe array ionization meter the variation multiple and have any different.For determining this difference, 1.5 times difference is preferred.That is, declaring gene before differentially expressed in treatment and untreated diseased cells, should find increases by 1.5 times or reduce 1.5 times at least at least through the cell for the treatment of is compared generation with untreated cell intensity.1.7 difference doubly is preferred, during genetic expression is measured 2 times or more times difference are most preferred.

A kind of method of the present invention relates to the gene expression profile of more various genes, to determine whether the people replys the use of therapeutical agent probably.Behind the gene expression profile of setting up respondent and non-responder's differentiation, each gene expression profile is fixed in the medium such as computer-readable medium, as mentioned below.Acquisition contains patient's sample of diseased cells (for example, for AML, the parent cell of green blood).Most preferably, this sample is the marrow sample, and extracts from patient's breastbone or iliac crest (iliac crest) according to ordinary method.Preferably, adopt ordinary method to carry out the marrow sucking-off, with the leukemic parent cell of enrichment.Then, obtain and amplification sample RNA from ill patient's cell, obtain the gene expression profile of the gene in the suitable gene bag, this preferably carries out (for big gene Bao Eryan) by microarray.Then with the express spectra of sample with analyzed the prognosis result before those compare.When a spot of gene was used in the gene bag, for example when using three gene profiles, simple nucleic acid amplification and detection scheme were the most preferred methods of measuring generegulation.In this case, can use other known amplification scheme of PCR, NASBA, rolling-circle replication, LCR and technician, wherein PCR is most preferred.When the gene bag comprised a large amount of genes or wish to measure multiple other expression of gene, it was preferred coming the evaluation form expression patterns according to the strength detection based on microarray mentioned above.

In a similar fashion, can carry out gene expression spectrum analysis reacts with monitor therapy.Aspect of this method, each stage during treating is applied to patient with the treatment of any appropriate therapies with gene expression analysis as indicated above.If gene expression pattern is consistent with positive result, then patient treatment is proceeded.If not so, then change treatment to the patient, as use other therapeutical agent, change dosage or cancel current treatment.This analysis makes can be before detectable clinical sign occurs, or in the face of otherwise intervene when being exactly ambiguous clinical sign and adjust treatment.

About molecular marked compound of the present invention, there are a lot of other form and methods to can be used for diagnostic uses.Methylating of genome area can influence gene expression dose.For example, but the supermethylation composing type ground down-regulation of gene expression of gene promoter region, and hypomethylation can cause the increase of steady-state mRNA level.Thereby, detect the alternative method that the methylate zone relevant with the gene of omen drug reaction, prognosis or state can be used as the diagnostic gene expression level.These class methods are well known to those skilled in the art.Additionally, the single nucleotide polymorphism (SNPs) that is present in promoter region also can influence the translation activity of gene.Therefore, detecting these SNPs by the known method of those skilled in the art also can be used as diagnosis and be used for detecting in difference and prejudge the differentially expressed gene of result.

Goods of the present invention are to can be used for treating, diagnose, predict, by stages and the otherwise representative of the gene expression profile of assess disease.Preferably, they are reduced to the medium that can read automatically, for example computer-readable media (magnetic, light, or the like).Described goods also can comprise the instruction of the gene expression profile that is used for assessing this media.For example, described goods can comprise CD ROM, and this CD ROM has the computer instruction of the gene expression profile that is used for comparison said gene bag.Described goods can also make gene expression profile be recorded in wherein with digital form, so that they can compare with the gene expression data from patient's sample.Perhaps, this express spectra can be with different manifestation records.Cluster (clustering) algorithm, from above-mentioned Partek, the swarm algorithm of " DISCOVERY " of Inc. and " INFER " software can be assisted visual these type of data well as incorporated.

Other goods of the present invention are the test kit that carries out said determination.Each this test kit preferably includes the specification sheets of people or machine-readable form and is used for the typical agents that described type is measured.These can for example comprise aforesaid, as to be configured to differentiate gene expression profile of the present invention nucleic acid array (for example cDNA or oligonucleotide array).They also can contain and are useful on the reagent that carries out nucleic acid amplification and detection, for example comprise reversed transcriptive enzyme, reversed transcriptive enzyme primer, corresponding PCR primer to, thermostable DNA polymerases such as Taq polysaccharase and the detection reagent that is fit to, such as but not limited to scorpion type (scorpion) probe, be used for probe, molecular beacon (beacon) probe, homogencous dyes primer that fluorescent probe measures or the fluorescence dye (as ethidium bromide) that is specific to double-stranded DNA.The test kit that is used to detect surface antigen contains coloring matter, or is based on antibody, the component that comprises for example for damping fluid, anti-antigenic antibody, detect enzyme and substrate, as the horse horseradish peroxidase or based on the reagent of vitamin H-avidin.The reagent constituents that is used to detect parent cell generally includes and is used to carry out flow cytometry method, parent cell and adheres to and measure and other common parent cell is measured reagent.

Conventional carcinostatic agent includes but not limited to tyrosine kinase inhibitor, MEK kinase inhibitor, P13K kinase inhibitor, map kinase inhibitor, apoptosis regulator and combination thereof.Wherein most preferred illustrative drug is " GLEEVEC " tyrosine kinase inhibitor, U-0126MAP kinase inhibitor, PD-098059MAP kinase inhibitor, SB-203580MAP kinase inhibitor and antisense nucleic acid, ribozyme and DNAzyme, Bcl-XL and the anti-apoptosis agent of Novartis.The example of the medicine that other is useful includes but not limited to United States Patent (USP) 6,306,897 calanolide; United States Patent (USP) 6,284, two lopps of 764 replacement; United States Patent (USP) 6,133,305 dihydroindolines; And the antisense oligonucleotide of United States Patent (USP) 6,271,210; Platinum complex such as cis-platinum or carboplatin, taxane compounds such as Paclitaxel or Docetaxel, camptothecine (camptothecin) compound such as Rinotecan (irinotecan) or Hycamtin (topotecan), antitumor vinca alkaloids such as vincaleucoblastine, vincristine(VCR) or vinorelbine (vinorelbine), antitumor nucleoside derivates such as 5 FU 5 fluorouracil, gemcitabine (gemcitabine) or capecitabine (capecitabine), mustargen or nitrosourea alkylating agent such as endoxan, Chlorambucil, carmustine, or lomustine, antitumor anthracycline derivative such as daunorubicin, Dx, or idarubicin; HER2 antibody such as trastzumab; And antitumor podophyllotoxin derivative such as Etoposide (etoposide) or teniposide (teniposide); And antiestrogen, comprise estrogen receptor antagon or selective estrogen receptor modulators, preferred Tamoxifen (tamoxifen), perhaps toremifene (droloxifene), droloxifene (droloxifene), faslodex and raloxifene (raloxifene), or aromatase inhibitor such as Exemestane (exemestane), Anastrozole (anastrozole), letrozole (letrazole) and vorozole (vorozole).

Carcinostatic agent also can comprise and relates to gene therapy or antisense therapy or RNA interferential therapeutical agent.These include but not limited to that sequence is complementary to the oligonucleotide of mRNA sequence, and it can be introduced into the translation of cell with blocking-up mRNA, thus the function of the gene of this mRNA of blocking-up coding.Use the oligonucleotide blocking gene to express and for example be described in Strachan and Read, HumanMolecular Genetics is in 1996.These antisense molecules can be the stable derivatives of the stable derivatives of DNA, DNA such as sulfo-phosphide or methylphosphonate, RNA, RNA as 2 '-O-alkyl RNA or other antisense oligonucleotide stand-in.Antisense molecule can be by microinjection, liposomes enclose or by being introduced into cell from the vector expression that carries antisense sequences.

In gene therapy, goal gene can be connected into virus vector, and they mediate treatment DNA transfer by infecting the acceptor host cell.The virus vector that is fit to comprises retrovirus, adenovirus, adeno-associated virus (AAV), simplexvirus, vaccinia virus, the scorching virus of marrow cinereum matter or the like.Perhaps, can will treat DNA by non-virus technology and shift into that cell is used for gene therapy, these technology comprise that employing part-DNA coupling or the receptor-mediated target DNA of adenovirus-part-DNA link coupled shift, the lipofection film merges or direct microinjection.These operations and variation thereof are suitable for exsomatizing and the vivo gene treatment.The molecular method scheme that is applicable to the gene therapy of gene is described in Gene Therapy Protocols, edited by Paul D.Robbins, Human press, Totowa NJ, 1996.

Can use separately with suitable dosage according to method compounds identified disclosed herein, this dosage is determined by routine test, so that obtain best inhibition or active minimized any genotoxic potential simultaneously.In addition, may want with common administration of other medicament or order administration.

The present invention further illustrates by following infinite embodiment.All documents that this paper quotes are all incorporated this paper by reference into.

Embodiment 1

The definition of clinical appraisal and reaction

Current research is the part of open label, polycentric, non-comparative 2 phases clinical study, the patient (Harousseau et al. (2003)) who wherein suffers from recurrence or refractory AML treats with Zarnestra, for initial continuous 21 days of each 28 day cycle, initial oral dosage is 600mg, twice of every day.The patient is divided into two groups: suffer from and recur AML's and suffer from refractory AML's.Treat 252 patients (135 recurrences with 117 refractories) altogether.Select 80 patients to provide the marrow sample to be used for the RNA microarray analysis, this is obtained individually the patient and agree.Total response rate is relatively low in this research.

Therefore, for the purpose of gene expression profile, to replying of Zarnestra be defined as the patient have objective response (alleviate [CR] fully, have the alleviation fully [CRp] of incomplete platelet recovery or partly alleviate [PR]), by the center observe or by the hematology that clinical scene is determined reply (the leukemia parent cell reduces greater than 50% in the marrow), by the center is observed and clinical scene is determined stable disease (not having progression of disease) but there is blood blood to reply.Alleviation fully with incomplete platelet recovery is defined similarly, only except the situation of platelet count less than 100,000/ μ L, is enough to guarantee blood transfusion independence.The part alleviation is defined as haematogonium at least 50% decline, and neutrophil leucocyte (＞500/ μ L) and platelet count (＞50,000/ μ L) with part are recovered.Replying to provide record to be confirmed after at least 4 weeks for the first time.

Sampling and microarray are handled

From gathering the marrow sample with the patient before the Zarnestra treatment, with phosphate buffered saline (PBS) (PBS) dilution, and use Ficoll-3, two (kharophen)-2,4 of 5-, 6-Triiodobenzoic acid salt (1.077g/mL) is centrifugal.With PBS washing white corpuscle twice, it is resuspended in the foetal calf serum (FBS) that contains 10% dimethyl sulfoxide (DMSO) (DMSO), and immediately-80 ℃ of preservations.Cell and adopt RNeasy Kit (Qiagen, Valencia CA) extract total RNA from cell sample thaws.Adopt Agilent Bioanalyzer to check the RNA quality.(Santa Clara, CA) scheme is carried out the synthetic of cDNA and cRNA according to Affymetrix.

Microarray is handled

If adopt one to take turns amplification, then for several samples, RNA output is too low and cRNA that can not obtain enough marks is used for chip hybridization, so carry out the two-wheeled linear amplification.For hybridizing, by at 40mM Tris-acetate, pH 8.1, hatch 35 minutes at 94 ℃ in 100mM potassium acetate and the 30mM magnesium acetate, to the cRNA random fragmentation of 11 μ g.Be set at the cRNA that makes fragmentation under 45 ℃ in the rotary baking box of 60rpm and U133A hybridization array 16 hours.After the hybridization, (with 6x SSPE and contain Triton X-100[0.005%] 0.5x SSPE) the washing array, and with streptavidin-phycoerythrin (SAPE that dyes; Molecular Probes, Eugene, OR).Quantitative employing Agilent G2500A GeneArray scanner to the probe of bonded mark carry out (Agilent Technologies, PaloAlto, CA).

The total fluorescence intensity of each array is normalized into unified value 600.Chip performance comes quantitatively by calculating signal/noise ratio (original signal/noise mean value)., then it is got rid of in further analyzing less than 5 as the fruit chip signal to noise ratio.In at least 10% chip, be regarded as " existence " as fruit gene, then they comprised in the into further analysis.According to these boundaries, remain 11,723 kinds of Affymetrix probe groups.By identify outlier and the normal distribution by analyzing gene intensity, the quality (Partek Pro V5.1) of coming further controlling gene expression data based on principle component analysis.

Statistical study

In order to identify the gene of replying, adopt the hundredths analysis with the hypersensitivity prediction.Identify and 40% non-responder contrast the gene that in 100% respondent, raises or reduce at least.Then chi square test and Student ' s t-check be used to that test patient is replied and patient's co-variation amount (comprising ras mutation status and genetic expression) between relevant significance.In Omniviz, carry out unsupervised k average and hierarchical clustering.Subsequently by staying one and stay five cross validation methods to analyze the predictive value of selected gene.Here, a kind of (or five kinds) sample is removed from data set, from 11,723 genes selectable marker once more.Then, adopt the predictive value of linear differential analysis to these genes of sample test of staying.Susceptibility is calculated as: the true positives number of detection adds false-negative summation divided by true positives.Specificity is calculated as: the true negative number of detection adds false-negative summation divided by true negative.Positive predictive value is calculated as: the true positives number is divided by true positives and false-positive number.Negative predictor is calculated as: the true negative number is divided by true negative and false-negative number.The positive likelihood ratio (likelihood ratio) of replying the patient of treatment deducts specificity for susceptibility divided by 1.Experimenter's characteristic working curve (ROC) is utilized for each sorter and selects suitable threshold, requires susceptibility 100%.The ROC diagnosis is each calculation of parameter susceptibility and specificity.

The real-time RT-PCR checking

The TaqMan real-time RT-PCR is used to check the microarray results of AHR and AKAP13 gene.For the RNA sample of each 1 μ g amplification, adopt T7 widow's (dT) primer and Superscript II reversed transcriptive enzyme to generate cDNA according to the explanation (Invitrogen) of manufacturers.Being used for the primer of AKAP13 gene and crt gene and MGB probe is Primer Express (Applied Biosystems) design, and be used for AHR gene and crt gene HPRT those can be used as Assays-on-Demand and obtain from ABI.Primer/the probe sequence that is used for AKAP13 is as follows: AKAP13 forward, 5 ' ggtcagatgtttgccaaggaa3 ' (SEQ ID NO:1); AKAP13 is reverse, 5 ' tcttcagaaacacactcccatc-3 ' (SEQ ID NO:2); The AKAP13 probe, 6FAM-tgaaacggaagaagcttgtA-3 ' (SEQ ID NO:3).

The all best after tested amplification efficiency of all primers and probe is more than 90%.Relative standard's curve is formed (in most of the cases, from 25ng to 2.5pg) by the HeLa cDNA of 5 extent of dilution (each 10 times).RT-PCR amplification mixture (25 μ L) contains 100ng template cDNA, 2x

Universal PC R mixture (master mix) (12.5 μ L; AppliedBiosystems), forward and reverse primer of 500nM and 250nM probe.Be reflected on the ABI PRISM7900HT sequential detector (Applied Biosystems) and carry out.Cycling condition is: at 50 ℃ of activation AmpErase UNG 2min, and at 95 ℃ of activated polymerization enzyme 10min, and 95 ℃ of 15sec of 50 cycle numbers and annealing temperature (59 ℃ or 60 ℃) 60sec.In each is measured, for goal gene and crt gene, typical curve and do not have the template contrast and included together with template cDNA, triplicate.The relative quantity of each gene is calculated based on typical curve, and proofreaies and correct with the amount of crt gene.The median variation coefficient (based on calculated amount) of three duplicate samples is 8%.Dependency between the template that employing is independently diluted from storing solution reruns is greater than 0.95.If double TaqMan experiment has shown reproducible results, then sample is only compared with microarray.

Clone is cultivated and AKAP13 crosses the mensuration of expression

AKAP13 carrier, oncoLBC and protoLBC and vehicle Control (pSRalpha-neo) obtain from Dr.Deniz Toksoz.Zheng et al. (1995).HL60 clone obtains from U.S. tissue culture collection institute (American Tissue Culture Collection), and it is incubated among the RPMI 1640 that contains 10%FBS.According to manufacturer specification, adopt Effectene reagent (Qiagen) with each carrier transient transfection cell, and placed 7 days down at G418 (600ug/mL).Then, (0,1.5,3.1,6.3,13,25,50,100,200,1000 and 10,000nM) adding is so that double cultivates (1.5 * 10 with various concentration with Zarnestra ⁵Cell/mL).At the 6th day pair cell counting.Cell counting is normalized to the culture that does not have medicine, to provide the per-cent of the control cells of living relatively.

The result

The express spectra of recurrence and refractory AML.

FTI is designed to suppress specifically the FTase activity at first, thus blocking-up oncogene ras approach.Therefore, we at first from 80 marrow DNA analysis of suffering from recurrence or the patient of refractory AML activation ras sudden change, and studied the ras sudden change and to the possible dependency between the replying of Zarnestra.Although 26% AML sample carries the N-ras sudden change, mutation status is not relevant with objective response or overall survival rate.Harousseau et al. (2003). therefore, we have carried out gene expression spectrum analysis and can be used for predicting the new mark that the FTI Zarnestra is replied with evaluation.Before with the Zarnestra treatment, obtain the marrow sample from 80 patients and be used for gene expression analysis.Table 1 has shown patient information.

Table 1. patient information

^*The stable disease (SD) of after independent studies person confirms, just being included only.

The HI=hematology is improved

CR=replys fully

PR=partly replys

The NR=no response

58 parts in 80 duplicate samples have been experienced quality control survey, have comprised RNA quality and chip performance.Between all the other patients of these 58 patients and this clinical study colony, produce risk factor, baseline values parent cell counting at age, sex AML kind (recurrence or refractory), cell, reply and the overall survival rate aspect do not have marked difference.

Table 2

Gene expression data and clinical information are integrated, and carried out retrospective analysis to identify the gene that can respondent and non-responder be separated with high-level susceptibility.These data are several screening steps of experience before differentiating differentially expressed gene.At first, remove the gene of at least 10% sample, not expressing.This reduces to 11,723 genes with number gene from about 22,000.Do not analyze for there being supervision, also get rid of in data centralization and show expression unconverted substantially gene (spreading all over all samples variation coefficient＜45%), and the percentage point stdn is used for remaining 5,728 genes.In this stage, carry out unsupervised k mean cluster analysis, so that identify any difference between them based on patient's whole gene expression profile.Six the main patient groups that adopted this technical evaluation.Between respondent and non-responder, do not observe breach (Fig. 1).Have only the minority gene may be relevant with the antitumous effect of FTIs, for example, the differential expression that might relate to the biological individual gene of FTI influences clinical response, and this may be covered by the noise that other 11,722 gene is introduced.

Embodiment 2

Discriminating differentially expressed gene between respondent and non-responder

Secondly, we adopt gene expression data to carry out supervision and analyze, to identify differentially expressed gene between all respondents and at least 40% non-responder.Can be selected these standards, so that identify the measurable gene of replying Zarnestra with the susceptibility of possible highest level.From 11,723 genes, identify except altogether 19 can and in the t-test, provide the gene of significant P value, (more details are referring to table 3 and table 10) with respondent and non-responder's classification.These genes comprise and relate to that signal transduction, apoptosis, cell proliferation, tumour take place and FTI biology (ARHH, AKAP13, those genes IL3RA) possibly.

19 oligogenes that table 3 prediction is replied Zarnestra and analytical results

The real-time RT-PCR conclusive evidence of gene marker

In order to examine the microarray gene expression data, cDNA is carried out the TaqMan real-time RT-PCR, wherein this cDNA is used to produce the target cRNA of the mark that is used for microarray hybridization.Selected two genes to be used to examine gene expression data.Select AHR and AKAP13 gene to be because these gene pairss respondent produces the specificity of highest level.For AHR, relation conefficient is 0.74, and for AKAP13, relation conefficient is 0.94, and this shows and can confirm microarray data (Fig. 2) by PCR.

Evaluation is as the AKAP13 gene of drug-resistance marker's thing

AKAP13 was expression among the patient that Zarnestra is failed to respond to any medical treatment.Adopt leaving-one method cross validation (LOOCV; Fig. 3 A) 58 samples has been calculated the predictor of this gene.AKAP13 genetic expression has been predicted no response with 96% negative predictive value (NPV), and low expression level is with 43% positive predictive value (PPV) (χ 2=13.7; P=0.0022) mediated and replied.Overall diagnosis accuracy is 69%, and the positive likelihood ratio of replying is 2.4.Therefore, based on the classificationization of AKAP13 genetic expression to this patient colony, with response rate from whole group 24% (14/58) increase in the low patient of expression of this gene 43% (13/30).The expression values of AKAP13 gene in each patient is presented among Fig. 3 B.When analyzing survival rate by the Kaplan-Meier analytical method, the low patient's who expresses of this gene intermediate value survival rate is 90 days, than the long (P=0.008 of the patient with high expression level; Fig. 3 C).

Identify the minimal set of 3 gene markers

LOOCV is used to the Candidate Set of identified gene mark, and these gene markers can be predicted replying Zarnestra with the accuracy higher than independent AKAP13.Set up sorter based on t check P value with the gene that increases number, and the error rate that adopts LOOCV to calculate this sorter keeps simultaneously predicting that the susceptibility of replying is at 100% (Fig. 4 A).

3 gene sorters can be replied (Fig. 4 A) with minimum error rate prediction.Also observe this point when staying five method cross validations.When adding more polygene, error rate increases, and shows that episome introduced noise to this sorter.For this 3 gene separating method, LOOCV shows: overall diagnosis accuracy be 74% and positive likelihood ratio be to have 94% NPV and 48% PPV at 2.9 o'clock.The associating expression values of this 3 gene is presented among Fig. 4 C among each patient.Therefore, concerning patient group, be 48% (12/25) to the response rate of Zarnestra, and be 24% (14/58) in this patient group with this genetic marker.

Adopt this 3 genetic marker (AHR, AKAP13 and MINA53), Kaplan-Meier analyzes once more and shows: the significant difference (Fig. 4 D) that has survival rate between respondent group who predicts and non-responder group.Compare with 31 patients that correctly are categorized as the non-responder, 13 patients that are categorized as the respondent improperly have better overall survival rate (Fig. 5).What is interesting is that 2 patients that mistake is divided into the non-responder only show the hematology improvement, and the overall survival time is relatively lacked (71,87 days).

AKAP13 cross to express increases among the AML resistance to Zarnestra.

The AKAP13 gene is to the most reliable mark of Zarnestra resistance.Therefore, we cross variant oncoLBC and the protoLBC that expresses this gene in HL60 clone, study its participation in FTI biology.Test the susceptibility of transient transfection then to Zarnestra.In this AML clone model, compare with control cells, cross the expressing of two kinds of AKAP13 variants cause near 20 times to the drug-fast increase of Zarnestra (Fig. 3).Curve is parallel compared with the control to move to right greater than 1 log unit from killing, and LBC oncogene and proto oncogene all increase resistance to Zarnestra with identical degree.

Discuss

Two groups have identified the gene expression profile of the adult AML of new diagnosis recently, and they can be used for predicting clinical effectiveness.Bullinger?et?al.(2004)；and?Valk?et?al.(2004)。These express spectras it seems than current use prejudge mark such as caryogram is more effective.In addition, predict that the express spectra that anticancer compound (comprising standard chemotherapeutics and new selectivity carcinostatic agent) is replied is found (Chang et al. (2003); Okutsu et al. (2002); And Cheok et al. (2003)).Hofmann?et?al.(2002)；and?McLean?et?al.(2004)。Similarly, with the patient the relevant pharmacogenetics spectrum of replying of tyrosine kinase inhibitor Gefitinib is found recently.Paez et al. (2004) and Lynch et al. (2004).In this research, the nonsmall-cell lung cancer patient subgroups has activation sudden change in the target EGF-R ELISA, and this sudden change is relevant with clinical response to this targeted therapy.

In the 2 phases research to the recurrence and the AML patient of refractory, we have identified the gene expression profile of predicting new farnesyl transferase inhibitor Zarnestra of replying.This compounds is just demonstrating at treatment haematological malignancies (Karp et al. (2001); Kurzrock et al. (2004); Alsina et al. (2003); Cortes et al. (2003); And Thomas et al. (2001)) and the prospect in the neurospongioma of noumenal tumour such as mammary cancer (Johnston et al. (2003)) and recurrent.Brunner?et?al.(2003)。Yet, although clinical response show, need day by day to be tested and appraised that most probable is replied medicine and therefore be that patient for the best experimenter of treatment customizes treatment.In addition, although ras is considered to the main target of this class medicine, several clinical studyes have confirmed that they may not be in the colony with high frequency ras sudden change effectively.Van Cutsem et al. (2004); And Rao et al. (2004).

Several gene markers of replying that might predict Zarnestra have been identified.These mark subclass are all predicted drug responses, and also think and might relate to FTI biology.One of best candidate thing of discovering from microarray is lymphocyte acute change (lymphoid blastcrisis) oncogene (oncoLBC or AKAP13).This gene plays Rho albumen guanylic acid exchange factor (Zheng et al. (1995)) and protein kinase A anchorin.Carr?et?al.(1991)。AKAP13 contains with the zone that comes from the helicoidal structure territory, and this helicoidal structure territory is known to interact with lamin B.Foisner?et?al.(1991)。This combination can cause lamin B to activate by protein kinase A, therefore increases mitogen activation.RhoB and lamin B are farnesylations, and are candidate's targets of FTIs.AKAP13 or proto-oncogene are because its 3 ' terminal disappearance causes cell transformation.Sterpetti?et?al.(1999)。Although it is to identify from the patient who suffers from chronic myelocytic leukemia at first, its expression is not recorded among the AML.

To several such viewpoints of evaluation support that may relate to the biological gene of FTI (ARHH, AKAP13IL3RA), i.e. the interaction of a plurality of approach can influence the function (Fig. 7) of FTIs in this AML patient group.These genes and several farnesylation protein-interactings that comprise ras, rho and possible lamin B.Rho albumen is the possible important antitumorgienesis target of FTIs.Sahai et al. (2002); With Lancet et al. (2003).RhoB, RhoA and RhoC have been considered to cross in the broad variety cancer and have expressed.Sahai?et?al.(2002)。In addition, RhoH (ARHH) usually resets in the tumour of spinal cord origin, and this rearrangement can cause it to cross expression.Pasqualucci?et?al.(2001)。Although most these Rho albumen are imperial ox Ji Longniu baseizations (geranygeranylated), closely interact mutually and with ras, RhoE and the little GTPase of RhoB of farnesylation between them.Sahai et al. (2002) and Li et al. (2002).In addition, confirmed that RhoH, RhoB and RhoE can act on the conversion capability of RhoA and RhoG in the antagonism mode.Li?et?al.(2002)。Guanylic acid exchange factor lymphocyte sudden turn of events oncogene (AKAP13) increases the activity of RhoA, and the activity that may also have other relevant little GTPase.Sterpetti et al. (1999); With Toksoz et al. (1994).In addition, AKAP13 can activate the nuclear lamina protein B by protein kinase A increases mitogen activation.Foisner?et?al.(1991)。IL3 receptor activation ras approach, this also is known.Testa?et?al.(2004)。This, as shown in Figure 7, the minimizing that the activity of IL3RA and AKAP13 increases and RhoH expresses can cause the transformant feature that increases.This may make the leukemia parent cell can overcome the anti-tumorigenesis effect of FTIs by compensation approach.On the contrary, when IL3RA and the low expression of AKAP13, and the active increase of RhoH, FTIs may be more effective in these approach of blocking-up.

At last, we confirm: the IC50 that expresses HL60AML clone that crosses of AKAP13 (oncoLBC and protoLBC variant) increases about 20 times.This shows that it is drug-fast correlating markings thing that crossing of AKAP13 expressed, and it may be the useful alternative drug targets of patient that Zarnestra is failed to respond to any medical treatment simultaneously.

Generally speaking, our discovery makes it possible to develop the diagnostic assay based on genetic expression, is hopeful to reply the patient of Zarnestra with evaluation.This information can be used for improving the direct treatment to the patient group who is fit to.Adopt survival rate as golden standard, this genetic marker has been predicted replying for the treatment of, and this can not predict by adopting conventional clinical response standard.Perhaps, this has proposed a problem: be used to predict treatment is replied to FTI this genetic marker whether also have with the FTI treatment irrelevant prejudge value.The mark of in the new diagnosis AML patient of conventional chemotherapy treatment, having identified before therefore, we have estimated of prejudging.Bullinger?et?al.(2004)。Although this mark has shown the application in current patient group, our 3 genetic markers further carry out layering to these bad organizing with good prejudging, and hold itself out to be the prediction thing that FTIs is replied.

Embodiment 3

Analyze AML and prejudge genetic marker

This 3 genetic marker can be independent of medication therapy type prediction prognosis.For this being confirmed we have at first estimated recently at genes identified presentation markup in the new diagnosis AML patient of conventional chemotherapy treatment.Bullinger?et?al.(2004)。Adopt the cDNA array to define this mark, so we are at first with these genes and the probe coupling that is present on the Affymetrix gene chip.In 133 predictability genes that people such as Bullinger identify, 167 probe groups (corresponding to 103 unique genes) and Affymetrix U133A chip coupling.3 genes identifying in our present analysis are not present in people's such as Bullinger 133 list of genes.SEQID NOs: be presented in the table 4.Adopt this 167 probe groups to define two main patient's groups (Fig. 8 A) by hierarchical clustering.Kaplan-Meier analyze to show these cluster evident layers turn to patient with good and bad prognosis (Fig. 8 B, p=0.000003).Therefore, this 133 gene prognostic markers subclass of our data acknowledgement Bullinger et al. (2004) evaluation also can be used for recurrence and refractory patient group.Thereby this shows that the prognosis gene profile is reliable surprisingly in different microarray platforms and different AML kind.

Cluster by this prognosis genetic marker definition does not have significantly more and respondent.Yet when these Zarnestra 3 genetic markers were used to this good and bad prognosis group, the patient who replys Zarnestra was by further from two prognosis component layers (Fig. 8 C).Therefore, this 3 genetic marker is marked with independent utility to prejudging, and it is specific to the FTI treatment in this patient group.

Embodiment 4

The definition of clinical appraisal and reaction

Current research is open label, polycentric, non-comparative 2 phases clinical study, inhibiting validity of survey procedure farnesyl transferase enzyme and security, wherein Zarnestra in AML as single medicament administration, for initial 21 days of each 28 day cycle, initial oral dosage is 600mg, twice of every day.The experimenter is divided into two groups: suffer from and recur AML's and suffer from refractory AML's.Treat 252 patients (135 recurrences with 117 refractories) altogether.Purpose for gene expression profile, replying of Zarnestra is defined as: the patient has objective response (CR, CRp, or PR) (as mentioned above), or the patient shows confirmed stable disease, or by the center observe or follow-up period between the hematology blood determined of clinical scene whenever reply (the leukemia parent cell reduces＞50%).

Sampling and microarray are handled

Obtain all samples from the patient, these patients agree described processing and analysis.Gather the marrow sample with before the Zarnestra treatment from the patient, with the PBS dilution and use Ficoll-3,5-pair of (kharophen)-2,4,6-Triiodobenzoic acid salt (1.077g/mL) is centrifugal.With PBS washing white corpuscle twice, be resuspended among the FBS that contains 10% dimethyl sulfoxide (DMSO) (DMSO), and immediately-80 ℃ of preservations.The cell that thaws, and (Qiagen, Valencia CA) extract total RNA from cell sample to adopt RNeasy Kit.Adopt Agilent Bioanalyzer to check the RNA quality.(Santa Clara, CA) scheme is carried out the synthetic of cDNA and cRNA according to Affymetrix.If adopt one to take turns amplification, then for several samples, RNA output is too low and cRNA that can not obtain enough marks is used for chip hybridization, so carry out the two-wheeled linear amplification.

For hybridization, the cRNA of 11 μ g is come random fragmentation hatching 35min under 94 ℃ in 40mM Tris-acetate pH8.1,100mM potassium acetate and 30mM magnesium acetate.In being set at the rotary oven of 60rpm under 45 ℃, the cRNA of fragmentation and U133A hybridization array 16h.After the hybridization, (with 6x SSPE and contain Triton X-100[0.005%] 0.5x SSPE) the washing array, and dye with streptavidin-phycoerythrin.Quantitative employing Agilent G2500A GeneArray scanner to the probe of bonded mark carry out (Agilent Technologies, Palo Alto, CA).

The total fluorescence intensity of each array is normalized into unified value 600.Come quantitative chip performance by calculating signal/noise ratio (original signal/noise mean value)., then it is got rid of in further analyzing less than 5 as the fruit chip signal to noise ratio.In at least 10% chip, be regarded as " existence " as fruit gene, then they be included in the further analysis.According to these boundaries, remain about 12, the 000Affymetrix probe groups.By identify outlier and the normal distribution (Partek Pro V5.1) by analyzing gene intensity, the quality of coming further controlling gene expression data based on principle component analysis.

Statistical study

In Omniviz, carry out unsupervised hierarchical clustering and cluster.Adopting S-Plus to carry out Kaplan-Meier analyzes.

Embodiment 5

The prognostic markers of identifying in newborn AML can be used for recurrence and refractory patient.

The adult AML patient's of new diagnosis gene expression profile described in two pieces of articles of delivering recently, and the purposes in the prediction clinical effectiveness.Bullinger et al. (2004); With Valk et al. (2004).We adopt Affymetrix U133A gene chip to analyze 58 express spectras of suffering from of suffering from recurrence and refractory AML.In 133 predictability genes that people such as Bullinger identify, 167 probe groups (corresponding to 103 unique genes) on the U133A chip, have been identified.Bullinger?et?al.(2004)。This 167 probe groups is listed in the sequence list, and specified SEQ ID NOs is presented in the table 4.

Adopt this 167 probe groups to define two main patient's groups (Figure 10 A) by hierarchical clustering.Kaplan-Meier analyze to show these clusters obviously be divided into patient with good and poor prognosis (Figure 10 B, p=0.0000219).Therefore, 103 subclass of this 133 gene prognostic markers of our data acknowledgement Bullinger et al. (2004) evaluation also can be used for recurrence and refractory patient group.Table 4.Thereby this shows prejudges gene profile in different microarray platforms, different AML kind and reliable surprisingly for different treatment algorithms.

Table 4

SEQ?ID?NO	Cluster 1 average	Cluster	2 averages	Ratio	The good prognosis group
						44	2.101613636	0.980102944	2.144278466	Raise
51	1.574545244	0.899302214	1.750852182	Raise
					52	2.459568465	1.161727897	2.117163987	Raise
55	1.752902097	0.968494897	1.809923937	Raise
					56	2.395730325	0.72630844	3.298502667	Raise
68	1.319861385	1.36567127	0.96645614	Downward modulation
					76	1.269874887	1.430330507	0.8878192	Downward modulation
90	0.986377684	1.432950683	0.688354244	Downward modulation
					91	1.177005842	1.428309412	0.824055231	Downward modulation
93	1.095239589	0.99028255	1.105986962	Raise
					104	1.798024918	0.806182842	2.230294202	Raise
108	5.041411016	0.60238936	8.369024013	Raise
					111	1.159807609	1.639962342	0.707216001	Downward modulation
114	1.300310388	1.324114868	0.982022345	Downward modulation
					115	1.342190037	2.306688065	0.581868896	Downward modulation
116	1.320735142	1.367135334	0.966060279	Downward modulation
					118	3.891659096	1.14773151	3.390739962	Raise
119	1.519690498	1.219697115	1.245957278	Raise
					120	1.328167684	1.250925424	1.061748094	Raise
124	4.052024058	0.844109372	4.800354307	Raise
					128	2.091653255	0.964077201	2.169591038	Raise
129	1.991638259	1.045332875	1.905267027	Raise
					130	2.231280889	0.889145566	2.509466362	Raise
131	1.815199475	0.992602441	1.828727596	Raise
					142	1.3198052	1.283492456	1.028292136	Raise
143	2.222738653	0.801693589	2.772553857	Raise
					144	1.591280353	1.037860182	1.533231913	Raise
147	1.050088807	1.267434876	0.828515	Downward modulation
					148	1.07046517	1.621321185	0.660242511	Downward modulation

150	1.768390061	0.83169323	2.126252803	Raise
					153	1.253048826	1.17309932	1.068152376	Raise
157	1.24240959	1.057597528	1.174747063	Raise
					158	1.039468561	1.565715718	0.663893546	Downward modulation
159	0.897290104	2.462204436	0.364425509	Downward modulation
					160	2.042794407	1.039120916	1.965887103	Raise
161	2.935364557	1.109613717	2.645393178	Raise
					167	0.771931017	1.585630652	0.486829021	Downward modulation
174	1.206470925	1.279664724	0.942802363	Downward modulation
					200	1.099482264	1.491611294	0.737110445	Downward modulation
207	4.901329448	0.871582063	5.623485909	Raise
					208	2.346190152	0.930016604	2.522740069	Raise
211	0.774893647	2.117870914	0.365883323	Downward modulation
					212	8.626480573	0.518118436	16.64963061	Raise
220	2.002466652	2.360710711	0.84824737	Downward modulation
					221	1.031573096	1.410790966	0.731201943	Downward modulation
222	1.236611762	1.579356606	0.782984512	Downward modulation
					224	4.252841813	1.038446727	4.095387566	Raise
227	2.330630017	0.740455045	3.147564506	Raise
					229	1.271065192	1.275443314	0.996567372	Downward modulation
230	4.079291876	0.770236845	5.296152606	Raise
					231	1.613398862	1.053542946	1.531403032	Raise
234	2.462533188	0.694618451	3.54515948	Raise
					235	1.736866874	1.312450625	1.323376926	Raise
236	0.973964566	1.442647881	0.675122862	Downward modulation
					237	2.146894817	1.117648799	1.920902898	Raise
238	1.121767102	1.496868758	0.749409122	Downward modulation
					241	1.09122088	1.120796083	0.973612325	Downward modulation
243	1.563910635	1.102945516	1.417940063	Raise
					244	1.473673649	1.474149259	0.999677367	Downward modulation
245	1.148794849	1.399237842	0.821014709	Downward modulation

246	1.350310245	1.327639719	1.017075812	Raise
					249	1.44906906	1.424150806	1.017496921	Raise
256	1.790625889	1.255091645	1.426689354	Raise
					260	1.423005429	1.45961841	0.97491606	Downward modulation
262	1.09603256	1.181485007	0.927673693	Downward modulation
					263	1.125580943	1.178287994	0.955268108	Downward modulation
264	0.891946685	1.753005455	0.508809988	Downward modulation
					265	1.2942793	2.776461422	0.466161456	Downward modulation
277	1.102290438	1.391074208	0.792402326	Downward modulation
					278	1.186998217	2.302906551	0.515434817	Downward modulation
281	0.79821098	1.919103099	0.415929181	Downward modulation
					288	1.236023874	1.187331666	1.041009778	Raise
301	1.087313678	1.212772631	0.896551959	Downward modulation
					303	1.820970134	1.219911387	1.492706891	Raise
306	0.893432913	1.529980396	0.583950563	Downward modulation
					310	1.076214323	1.702252048	0.632229713	Downward modulation
314	1.407137316	1.229815501	1.144185705	Raise
					315	2.943883289	0.906317573	3.248180746	Raise
316	1.666394606	0.87032571	1.914679281	Raise
					340	1.499937462	0.921725287	1.627315083	Raise
346	1.035229803	1.508152041	0.686422705	Downward modulation
					347	1.317050838	1.350558887	0.975189495	Downward modulation
348	1.481608791	1.161107338	1.276030856	Raise
					351	1.177285398	1.293627618	0.910065139	Downward modulation
352	1.328716012	1.354505535	0.980960194	Downward modulation
					353	1.245636653	1.388354047	0.897203891	Downward modulation
354	1.057630522	1.407147783	0.751612968	Downward modulation
					355	1.394737541	1.22633541	1.137321429	Raise
356	1.194410314	1.007530697	1.185482803	Raise
					359	1.653993358	1.201194991	1.37695659	Raise
361	1.318561503	1.233384342	1.069059707	Raise

364	1.045812959	1.463795973	0.714452682	Downward modulation
					365	1.721871636	1.213128563	1.419364517	Raise
366	1.437978693	1.211203408	1.18723138	Raise
					369	1.480402866	1.289306967	1.148215983	Raise
373	1.750186104	1.199777316	1.458759122	Raise
					379	1.987020453	2.331667877	0.852188458	Downward modulation
383	1.419274906	1.587359923	0.894110331	Downward modulation
					385	1.412275401	1.410313065	1.001391419	Raise
387	1.339345508	1.331753924	1.005700441	Raise
					395	1.747595879	1.473302004	1.186176272	Raise
401	1.237925661	1.240617377	0.997830341	Downward modulation
					404	1.176676928	1.14697739	1.025893743	Raise
407	1.256304659	1.357899094	0.925182633	Downward modulation
					415	2.013680769	0.801461983	2.512509405	Raise
418	1.405740421	1.158990553	1.212900673	Raise
					423	1.346444485	1.136343796	1.184891835	Raise
426	9.592759013	1.560596128	6.146855576	Raise
					430	1.768393033	1.23257422	1.434715252	Raise
437	1.40453464	1.860231401	0.755032218	Downward modulation
					438	1.673447728	3.930927905	0.425713157	Downward modulation
445	1.283792446	1.501667999	0.85491097	Downward modulation
					446	1.214492279	1.091382628	1.11280155	Raise
448	1.561740275	1.228333636	1.271430033	Raise
					450	1.734251715	1.844252606	0.940354759	Downward modulation
452	1.408580649	1.582858413	0.889896807	Downward modulation
					453	1.1383915	1.741970797	0.653507798	Downward modulation
454	1.079716972	1.450127515	0.744566917	Downward modulation
					457	1.783529874	1.314108612	1.357216487	Raise
460	1.136773942	1.003606776	1.132688588	Raise
					462	1.220645614	1.113413039	1.096309789	Raise
466	1.183017496	1.211183566	0.976745003	Downward modulation

467	1.113306085	1.067417158	1.042990621	Raise
					469	1.211346488	1.1871952	1.020343148	Raise
484	0.998901987	1.654269584	0.60383265	Downward modulation
					485	1.133736575	1.914231031	0.592267368	Downward modulation
489	1.412382921	1.384849519	1.019881873	Raise
					493	3.988440713	0.965026485	4.132985753	Raise
495	1.187128423	1.462785203	0.81155348	Downward modulation
					498	3.691405839	0.765303503	4.823453473	Raise
499	2.216130138	0.848821568	2.610831558	Raise
					507	2.199861346	0.773086467	2.845556663	Raise
520	1.67633492	1.194585675	1.403277265	Raise
					524	7.743976211	0.841225224	9.205592025	Raise
529	2.591731743	1.08342825	2.392158173	Raise
					530	1.273363236	1.097292728	1.160459013	Raise
535	1.095879149	1.302490014	0.8413724	Downward modulation
					546	0.987046127	2.124625445	0.464574181	Downward modulation
550	2.103990301	0.929475745	2.263631205	Raise
					555	1.439273076	1.211738853	1.187774967	Raise
557	1.121983318	1.251788385	0.896304305	Downward modulation
					559	1.197910138	1.34693432	0.889360468	Downward modulation
560	1.144102989	1.261720239	0.906780246	Downward modulation
					565	1.078544278	1.162115183	0.928087244	Downward modulation
568	1.46401688	0.960152377	1.524775561	Raise
					569	1.262437604	1.105379077	1.142085669	Raise
585	1.349621283	1.10729713	1.218842935	Raise
					590	1.203037349	1.068047506	1.126389361	Raise
609	1.395352279	1.172354292	1.190213819	Raise
					624	1.131210862	1.399636303	0.80821772	Downward modulation
630	0.859929427	1.37477642	0.625504929	Downward modulation
					637	0.72293765	2.343345902	0.308506588	Downward modulation
641	1.340847351	1.601484153	0.837252962	Downward modulation

?656	1.259818484	1.213892186	1.037833918	Raise
					?658	0.60640428	2.092078796	0.289857285	Downward modulation
?659	1.099549227	1.166865002	0.942310572	Downward modulation
					?665	1.615399946	1.733324276	0.931966378	Downward modulation
?684	0.734401434	2.496195783	0.294208267	Downward modulation
					?685	2.158835355	0.765517273	2.82010012	Raise
?686	0.957971846	1.386779118	0.690789061	Downward modulation
					?687	0.982131268	1.162818279	0.844612856	Downward modulation
?690	2.421692274	0.791473723	3.059725425	Raise
					?691	1.268020946	0.973474927	1.302571757	Raise
?692	1.054213523	1.147045848	0.91906834	Downward modulation
					?693	1.251020003	1.271391137	0.983977288	Downward modulation
?694	1.122443987	1.153661375	0.972940597	Downward modulation
					?696	1.301034813	1.029185966	1.264139675	Raise

This prognostic markers has nothing to do with 3 genetic markers that prediction is replied Zarnestra.

We have identified the genetic marker (AHR, AKAP13, MINA53) that prediction is replied Zarnestra in recurrence and refractory AML patient.These genes the patient can be categorized as have good and poor prognosis as a result group (Figure 11 B, p=0.002).Produced such query: this genetic marker is to predict replying that FTI treats, and still only identifies the patient with general good prognosis.When this 3 genetic marker is used to good and during the poor prognosis group, the respondent by further from this prognosis group categories (Figure 11 C, p=0.000003).After adopting these two kinds of genetic markers, Zarnestra is not produced the patient group who replys and have a poor prognosis tangible classification, this with the treatment type irrelevant (Figure 11 D, p=0.0000005).Therefore as if, this 3 genetic marker is independent of the mark of having identified of prejudging, and it is specific to the FTI treatment in this patient group.Thereby we think that this prognostic markers perhaps can unite use with drug specificity mark (as Zarnestra prediction spectrum), are used for managing patient treatment better.

Embodiment 6

Evaluation is differentially expressed gene between respondent and non-responder's (not comprising the stable disease patient)

From this is analyzed, remove 4 patients, have stable disease because they are classified as, and these patients can not be defined as respondent or non-responder clearly.Comprise stable disease patient into may be used for selecting relevant and deviation occurs with prognosis with the analysis of the irrelevant gene of pharmacological agent.This causes 10 respondents and 44 non-responder's contrasts.Selected gene requires the specificity of demonstration 40% and 2.0 minimum average B configuration to change multiple.Can select following standard for use, described standard makes and can identify the gene that prediction can be replied Zarnestra with the susceptibility of possible highest level.Identified 8 genes altogether from 11,723 genes, these 8 genes can be classified by (table 5) to respondent and non-responder, and provide significant P value (P＜0.05) in the t check.These genes comprise and relate to signal transduction, apoptosis, cell proliferation, tumour generation and may relate to biological those genes of FTI.AKAP13 is the most reliable mark.

Then, we are devoted to identify the minimal set with the gene of the diagnosis accuracy that offers the best from these 8 selected genes.Based on the AUC value, set up sorter with the gene that increases number from the analysis of recipient's operating characteristic, and adopt LOOCV to calculate the error rate of these sorters, the susceptibility that keeps prediction to reply is that 100% (Figure 12 is a) simultaneously.AKAP13 gene sorter can reply that (Figure 12 a) less than the prediction of 40% lowest error.When being used for this sorter greater than 2 genes, error rate is increased to and surpasses 50%.For AKAP13, it is 63% that LOOCV is presented at overall diagnosis accuracy, and positive likelihood ratio is to have 93% NPV and 31% PPV (Figure 12 b) at 2.0 o'clock.The expression values of AKAP13 is presented among Figure 12 c among each patient.Therefore, concerning having patient's group that low AKAP13 expresses, be 31% (8/26) to the response rate of Zarnestra, and be 18% (10/54) in current patient group.Adopt the AKAP13 gene, Kaplan-Meier analyzes and shows that there is significant difference (Figure 12 d) in survival rate between respondent's group of predicting and non-responder's group.

Table 5 is predicted the tabulation of 8 oligogenes that Zarnestra is replied

?SEQ?ID?NO：	Symbol	AUC	Change multiple	The P value	Functional description
						309	AKAP13	0.830	0.491	0.00007	Signal in the cell, knurl takes place
151	AHR	0.807	0.446	0.00019	Signal transduction, apoptosis
						222	SCAP2	0.777	0.431	0.00007	Signal transduction
496	NPTX2	0.738	0.115	0.02934	Cell adhesion
						451	BAT1	0.725	0.458	0.00097	Cell biological is synthetic
272	IL3RA	0.705	0.375	0.00226	Receptor signal
						411	IDS	0.645	0.395	0.00069	Metabolism
280	P2RY14	0.627	0.369	0.00145	Signal transduction

The area under curve that AUC=analyzes from recipient's operating characteristic.This is the indication of overall diagnosis accuracy.

Embodiment 7

Can prediction suffer from the patient of new diagnosing acute myelomatosis to Zarnestra (

R115777) the gene expression profile of replying.

Zarnestra (

R115777) be proved in suffering from the patient of blood disease and have clinical response.Although the arrestin farnesylation is significant feature mechanism (MOA), farnesyl inhibition level is not the reliable pharmacodynamics mark of replying, and does not know that what genetic marker can be used to prediction and replys yet.Design this research likely and be intended to identify potential genetic marker and presentation markup, these genetic markers and presentation markup may substitute the predictor of replying Zarnestra in acute myelogenous leukemia (AML) patient.In the low risk patient of the AML that suffers from new diagnosis, collect marrow sample and analyzing gene express spectra from 2 phase of the single armed clinical study of Zarnestra.Lancet?et?al.(2004)。Altogether, before Zarnestra treatment (n=25), among (n=30) and afterwards (n=24) 79 samples have been carried out expression pattern analysis.

Adopt Affymetrix U133A Gene

Array analysis marrow sample.Total genetic expression mark discloses the variation that the Zarnestra treatment causes genetic expression, and they stop the back in treatment and kept as many as 120 days.Sample before and after farnesyl transferase suppress to be handled is compared, identify except about 500 at the gene that has noticeable change aspect the genetic expression (false discovery rate [FDR]＜0.005), comprise that several genes relevant with farnesylation are (as K-ras, FNTA).The gene of many adjustment be accredited as relate to obviously that protein biology is synthetic, the gene of dna replication dna, intracellular signal transduction and cell cycle approach, so inhibition of reaction pair cell proliferation.Also identified the subclass (comprising the gene relevant) of 27 genes of difference adjustment between respondent and non-responder (P＜0.01) with signal transduction and cell cycle.Also in the sample before processing to before from recurrence and 2 clinical trial phases of refractory AML the genes identified presentation markup test, predict the ability of replying to study their.Raponi?et?al.(2004)。6 assortments of genes are found in this independent sets of sample has significant prediction accuracy (P=0.05).Genes identified may be used as the active alternative biomarker of Zarnestra from these researchs.

Patient's sample

In the research of 2 phases, the patient who suffers from new diagnosis AML accepts

600mg bid21dQ4wks.

Before the treatment, among and obtain the marrow sample afterwards.By Ficoll-Hypaque density centrifugation separating monocytic cell, and freezing in the mode that can survive.

Microarray analysis

From patient's parent cell amplification messenger RNA(mRNA), and with Affymetrix U133A chip hybridization, detectable about 22,000 genes of this chip (Figure 13).Chip data is by pre-filtering, with the gene of removing low quality data and not expressing in patient's sample of at least 10%.In addition, remove the gene that does not have to change (CV＜40%) in data centralization.Remain about 8000 genes and be used for further analysis.The quality control of 79 chips experience is measured altogether, and has relevant clinical response data.

Statistical study

For each gene, check effect and time and their interaction of studying pharmacological agent with variance analysis (ANOVA) and t.By discovery rate (FDR) algorithm of application error, control multihypothesis test (multiple hypotheses testing).All statistical study are all carried out in S-Plus 6.1 (Insightful Corporation).In Partek Pro, carry out principle component analysis.Adopt relativity measurement and complete connection method to carry out hierarchical clustering (OmniVizPro ^TM, OmniViz, Maynard, MA).Adopt Gene Ontology functional classification to carry out path analysis.Table 6 has shown analytical results.

The functional gene classification that table 6. is significantly regulated by Zarnestra

GO.ID	GO.Class	The P value
			6886	The intracellular protein transhipment	5.75E-05
6951	Heat-shocked is replied	8.32E-05
			6913	Nucleo-cytoplasmic transport	1.18E-04
6207	Pyrimidine base is biosynthesizing from the beginning	1.58E-04
			6809	The nitrogen protoxide biosynthesizing	1.58E-04
6376	The mRNA splice site is selected	2.05E-04
			245	The spliceosome assembling	2.05E-04
6259	The DNA metabolism	4.86E-04
			6371	The mRNA montage	4.86E-04
6607	Carry the nuclear input of the substrate of NLS	5.64E-04
			15031	Protein transport	6.05E-04
6338	Chromatin Remodeling (remodeling)	9.87E-04
			6397	MRNA processing	0.003201
7050	Cell-cycle arrest	0.003685
			8380	The RNA montage	0.003992
6512	The ubiquitin circulation	0.004639
			6396	RNA processing	0.005096
398	Nuclear mRNA montage by spliceosome	0.007120
			6916	Anti-apoptosis	0.007395
6118	Electron transport	0.010004
			7049	Cell cycle	0.019631
6457	Protein folding	0.021817
			7264	Little GTPase Mediated Signal Transduction	0.025281
8285	The negative adjusting of cell proliferation	0.026482
			6366	From transcribing of PolI promotor	0.048613

About 8000 genes are used to total nothing supervision cluster.Sample before the sample distally clustering processing after the treatment neutralization treatment.Sample after the processing comprises that those stop 120 days patient's of back as many as sample from treatment.Figure 14 shows: stop back AML sample in the Zarnestra treatment and keep total changes in gene expression of FTI mediation.

Discovery is handled back 502 genes differentially expressed (p＜0.005) with Zarnestra.These genes are listed among the table 7A-7C, in more detail in table 9.

Be included in having in these tables and relate to FTI biology a lot of genes of (comprising AKT1, CENPF, KRASRAF1, STATs and farnesyl transferase).Certain several gene function type (table 8) is rich in discovery significantly in this gene table.

Differentially expressed between table 8. Zarnestra treatment back respondent and the non-responder

?SEQ?ID NO：	Probe groups ID	Gene function (GO)	?RE?pre?v?RE post	PD?pre?v?PD?post
					13	200044	The RNA montage	Raise	Indifference
23	200661	Albumen	Raise	Indifference
					85	201804	Protein folding	Raise	Indifference
123	202349	Protein folding	Raise	Indifference
					137	202622	Nuclear	Raise	Indifference
149	202761	Cytoskeleton	Raise	Indifference
					198	203845	Transcribe	Downward modulation	Indifference
210	204067	Electron transport	Raise	Indifference
					218	204215	Film	Downward modulation	Indifference
257	205339	Cell proliferation	Downward modulation	Indifference
					266	205644	The RNA montage	Downward modulation	Indifference
268	205807	Bone mineralization	Downward modulation	Indifference
					290	207163	Signal transduction	Raise	Indifference
329	208819	Signal transduction	Raise	Indifference
					336	208927	RNA	Raise	Indifference
362	209295	Signal transduction	Downward modulation	Indifference
					428	211762	Cell cycle	Downward modulation	Indifference
470	212833	Transhipment	Downward modulation	Downward modulation (higher in PD)
					521	214298	Cell cycle	Raise	Raise (higher in PC)
549	215764	Transhipment	Raise	Raise (higher in PD)
					551	215905	The RNA montage	Raise	Indifference
598	218256	Transhipment	Downward modulation	Indifference
					610	218373	Apoptosis	Raise	Indifference
619	218603	Cell cycle	Downward modulation	Indifference
					660	220671	Transcribe	Indifference	Downward modulation
698	44696	Signal transduction	Indifference	Raise
					699	49485	Transcribe	Raise	Indifference

We had identified 8 energy drug-fast gene of prediction Zarnestra in recurrence and refractory AML in the past.Gene in the tool predictability of this data centralization is AKAP-13 (AUC=0.83).

Newly diagnosing out the current concentrated of AML sample, the predictor (CTEP20) of testing these genes.Use is replied or the patient's of PD sample fully from having.The result is presented among Figure 15.

6 genes are set up the prediction sorter from the top, because these genes provide in training set less than 50% error rate.This 6 gene sorter is presented in the table 9, and it is presented among Figure 16 the ability that new diagnosis AML classifies.

Table 9

SEQ?ID?NO：	PSID	Symbol	INT17AUC	CTEP20AUC
					309	208325	AKAP13	0.830	0.464
151	202820	AHR	0.807	0.750
					222	204362	SCAP2	0.777	0.893
496	213479	NPTX2	0.738	0.766
					451	212384	BAT1	0.725	0.288
272	206148	IL3RA	0.705	0.714
					411	210666	IDS	0.645	0.446
280	206637	P2RY14	0.627	0.589

Conclusion

27% clinical marrow sample can be used, and illustrating needs to optimize sampling.

The total genetic expression mark of AML cell shows: the FTI treatment makes treatment stop the back and produces stable changes in gene expression.

About 500 genes (p＜0.005) are subjected to the effect of Zarnestra, and this reflects FTIs target number of ways.The many approach relevant before this comprises with FTI biology.

After the Zarnestra treatment, between respondent and non-responder, find the differential expression of 27 genes.These are candidate PD marks.

The 6 gene sorters of identifying in recurrence and refractory AML are found among the new diagnosis AML has predictive value.

Embodiment 8

Antibody (prophesy)

LBC oncogene derived peptide is synthesized, with the keyhole limpet hemocyanin coupling and be used for immunize rabbit to produce polyclonal antibody.With the reactivity of ELISA test sera to corresponding peptides, and positive batch of affinity purification.Antibody purification energy specific detection has epi-position in tissue slice peptide.If when this corresponding peptide and this antibody added simultaneously, signal was eliminated fully, then this is examined.Except that this polyclonal antibody that is suitable in IHC, generation can detect this proteic monoclonal antibody of natural folding.In order to generate monoclonal antibody, generate the antigen of purifying, this antigen generates in mammalian cell to guarantee natural folding and post transcriptional modificaiton.This antigen, LBC onco albumen-IgG constant portion fusion rotein is expressed in murine myeloma cell, adopts the Fc part as this albumen of bait (bait) purifying.The antigen of this purifying is discerned by the C-terminal polyclonal antibody in the Western trace.By select produce with the LBC reactive polypeptide and not with the positive colony of the antibody of IgG constant portion reaction, with this antigen with the mouse monoclonal antibody that generates anti-LBC peptide.Can adopt these and similar antibody easily to prepare the test kit that is used for clinical identification LBC oncogene.Such test kit comprise be used to identify the LBC peptide antibody (thus, the LBC oncogene), the indicator of Shi Heing (for example, enzyme, marker or the like) and (randomly) other reagent of the clinical application that can be used for this test kit, be generally used for the material of this type of mensuration as dilution buffer liquid, stablizer and other.

Embodiment 9

Immunohistochemistry (indication)

Will at the C-terminal peptide of LBC oncogene, be used for through the polyclonal antibody of protein affinity purification that IHC detects and location LBC oncogene.Formalin fixed and paraffin-embedded 4um section are placed on the slide glass of 3-aminopropyl-triethoxy-silicane (APES, Sigma, St.Louis MO) coating.With these section dewaxings, rehydration in the ethanol of gradient concentration, and at room temperature handled 30 minutes with methyl alcohol superoxide (0.5% hydrogen peroxide in the anhydrous methanol), to block endogenous peroxidase activity.In microwave oven, carry out twice antigen and reclaim each 5 minutes (650W).With Elite ABC test kit (Vectastain, Vector Laboratories, Burlingame CA) to immunoperoxidase staining.The LBC peptide antibody uses with the dilution in 1: 2000 of the best.The avidin mixture of biotinylated second antibody and peroxidase labelling was hatched 30 minutes in section.Dilution in PBS (pH 7.2), all is incubated in the damp camera, at room temperature carries out.Between different staining procedures, clean three slide glasss with PBS.This peroxidase stain at room temperature developed the color 15 minutes with 3-amino-9-ethyl carbazole (Sigma) solution (0.2mg/ml in the 0.05M acetate buffer solution wherein contains 0.03% hydrogen peroxide, and pH 5.0).At last, redye section a little with Mayer ' s phenodin, and with moisture mounting medium (Aquamount, BDH) mounting.In control experiment, this first antibody is replaced by the IgG component of normal rabbit serum or with this first antibody of LBC peptide preadsorption.These dyeing show that the LBC oncogene is present in the cell subsets.

For the clear purpose of understanding foregoing invention, by drawings and Examples it is had been described in detail, but these explanations and embodiment should not be counted as limitation of the scope of the invention.

Table 10. sequence table is described

SEQ ID?NO	psid	Describe	Title	Registration number
						1		The AKAP13 forward primer		NM_007209
2		The AKAP13 reverse primer		NM_001494
						3		The AKAP13 probe		NM_000975
4	200002	Ribosomal protein L 35	RPL35	NM_007209
						5	200008	The GDP inhibitor 2 that dissociates		NM_001494
6	200010	Ribosomal protein L 11		NM_000975
					7	200013	Ribosomal protein L 24	RPL24	NM_000986
8	200017	Ribosomal protein S27a	RPS27A	NM_002954
					9	200018	Ribosomal protein S13	RPS13	NM_001017
10	200025	Ribosomal protein L 27	RPL27	NM_000988
					11	200026	Ribosomal protein L 34	RPL34	NM_000995
12	200041	HLA-B associated retroviral-1	D6S81E	NM_004640
					13	200044	Splicing factor 9 is rich in arginine Serine (arginineserine)	SFRS9	NM_003769
14	200056	Highly similar with integrin alpha-7	FLJ12486	AK022548
					15	200061	Be similar to ribosomal protein S24		BC000523
16	200073	HnRNP-C sample albumen		M94630
					17	200086	Cytochrome c oxidase subunit IV		AA854966
18	200087	FLJ21323	FLJ21323	AK024976
					19	200091	Ribosomal protein S25		AA888388
20	200634	profilin?1	PFN1	NM_005022
					21	200640	The single methoxy enzyme activation of tyrosine 3-monooxygenase tryptophane 5-albumen, the ζ polypeptide	YWHAZ	NM_003406
22	200643	High-density lipoprotein (HDL) is conjugated protein	HDLBP	NM_005336
					23	200661	The protected protein of beta galactosidase enzyme	PPGB	NM_000308
24	200718	Transcriptional elongation factor B (SIII), polypeptide 1 sample		NM_003197
					25	200772	Prothymosin, α (gene order 28)		NM_002823

26	200780	Guanine nucleotide binding protein (G albumen), α stimulating activity polypeptide 1	GNAS1	NM_000516
					27	200801	The β Actin muscle	ACTB	NM_001101
28	200834	Ribosomal protein S21	RPS21	NM_001024
					29	200846	Protein phosphatase 1, catalytic subunit, α abnormal shape	PPP1CA	NM_002708
30	200853	The H2A histone family, member Z	H2AFZ	NM_002106
					31	200857	Nuclear receptor corepressor 1	NCOR1	NM_006311
32	200858	Ribosomal protein S8	RPS8	NM_001012
					33	200902	The 15kDa seleno-protein	SEP15	NM_004261
34	200925	Cytochrome c oxidase subunit Via polypeptide 1	COX6A1	NM_004373
					35	200934	DEK oncogene (DNA combination)	DEK	NM_003472
36	200949	Ribosomal protein S20	RPS20	NM_001023
					37	200964	Ubiquitin activating enzyme E1	UBE1	NM_003334
38	200971	The endoplasmic reticulum albumen 1 that stress be correlated with	SERP1	NM_014445
					39	200981	Neuroendocrine albumen 55	NESP55	NM_016592
40	200991	The KIAA0064 gene product	KIAA0064	NM_014748
					41	200999	Transmembrane protein (63kD), zone between the endoplasmic reticulum golgi body	P63	NM_006825
42	201019	Eukaryotic translation initiation factor 1A	EIF1A	NM_001412
					43	201027	The KIAA0741 gene product	IF2	NM_015904
44	201042	Trans-glutaminases 2		AL031651
					45	201049	Ribosomal protein S18	RPS18	NM_022551
46	201094	Ribosomal protein S29	RPS29	NM_001032
					47	201104	DJ328E19.C1.1		NM_015383
48	201118	PDG	PGD	NM_002631
					49	201134	Cytochrome c oxidase subunit VIIc	COX7C	NM_001867
50	201163	RhIGF-1 albumen 7	IGFBP7	NM_001553
					51	201195	L-amino acid transporter 1		NM_003486
52	201212	Proteolytic enzyme, halfcystine, 1	legumain	NM_005606
					53	201227	The inferior complex body of nadh dehydrogenase 1 β, 8	NDUFB8	NM_005004
54	201244	V-raf-1 murine leukemia virus oncogene homologue 1	RAF1	NM_002880

55	201249	Solute belongings (solute carrier) 2 members 1 of family	SLC2A1	NM_006516
					56	201250	2 members 1 of solute belongings family	SLC2A1	NM_006516
57	201273	Signal recognition particle 9kD	SRP9	NM_003133
					58	201277	Heterogeneous nuclear ribonucleoprotein AB	HNRPAB	NM_004499
59	201300	PrPC (p27-30)	PRNP	NM_000311
					60	201305	Be rich in leucic acidic protein		NM_006401
61	201317	Proteasome (prosome, macropain) sub α 2 types	PSMA2	NM_002787
					62	201324	Epithelial cell membranin 1	EMP1	NM_001423
63	201352	YME1 (S.cerevisiae)-sample 1	YME1L1	NM_014263
					64	201381	Calcyclin is conjugated protein		NM_014412
65	201393	RhIGF-1 2 acceptors	IGF2R	NM_000876
					66	201403	Microsome glutathione S-transferase 3	MGST3	NM_004528
67	201429	Ribosomal protein L 37a	RPL37A	NM_000998
					68	201445	Acid calponin 3	CNN3	NM_001839
69	201455	The responsive aminopeptidase of tetracycline		NM_006310
					70	201472	Von Hippel-Lindau conjugated protein 1	VBP1	NM_003372
71	201483	Ty (S.cerevisiae) repressor 4 homologues 1		NM_003168
					72	201568	The lower molecular weight ubiquinone is conjugated protein	QP-C	NM_014402
73	201588	Trx-sample, 32kD	TXNL	NM_004786
					74	201597	Cytochrome c oxidase subunit VIIa polypeptide 2	COX7A2	NM_001865
75	201620	Film is in conjunction with the transcription factor protein enzyme, site 1	MBTPS1	NM_003791
					76	201621	Neuroblastoma suppresses tumour and takes place 1	NBL1	NM_005380
77	201635	Fragile X amentia, euchromosome homologue 1		U25165
					78	201665	Ribosomal protein S17	RPS17	NM_001021
79	201675	A kinases (PRKA) anchorin 1	AKAP1	NM_003488
					80	201699	Proteasome 26S subunit ATP enzyme 6	PSMC6	NM_002806
81	201715	KIAA0670	KIAA0670	NM_014977
					82	201716	Sorting connects albumen 1	SNX1	NM_003099
83	201738	Translation factor sui1 homologue	GC20	NM_005875
					84	201754	Cytochrome c oxidase subunit VIc	COX6C	NM_004374

85	201804	Cytoskeleton related protein 1	CKAP1	NM_001281
					86	201805	Protein kinase, AMP-activatory, γ 1 on-catalytic subunit	PRKAG1	NM_002733
87	201812	6.2kd albumen	LOC54543	NM_019059
					88	201818	Putative protein FLJ12443	FLJ12443	NM_024830
89	201890	Ribonucleotide reductase M2 polypeptide		NM_001034
					90	201910	RhoGEF (ARHGEF) and pleckstrin domain protein 1	FARP1	NM_005766
91	201911	RhoGEF (ARHGEF) and pleckstrin domain protein 1	FARP1	NM_005766
					92	201921	Guanine nucleotide binding protein 10	GNG10	NM_004125
93	201934	Putative protein PRO2730		NM_025222
					94	201938	Disappearance in oral carcinoma 1	DOC1	NM_004642
95	201987	The thryoid receptor associated protein, the 240kD subunit		NM_005121
					96	202001	The inferior complex body of nadh dehydrogenase 1 α, 6	NDUFA6	NM_002490
97	202029	Ribosomal protein L 38	RPL38	NM_000999
					98	202077	Nadh dehydrogenase 1, the inferior complex body of α β, 1	NDUFAB1	NM_005003
99	202078	COP9 subunit 3	COPS3	NM_003653
					100	202090	Ubiquinol-cytochrome c reductase (6.4kD) subunit	UQCR	NM_006830
101	202110	Cytochrome c oxidase subunit VIIb	COX7B	NM_001866
					102	202114	Sorting connects albumen 2	SNX2	NM_003100
103	202141	The COP9 homologue		NM_006710
					104	202154	Tubulin, β, 4	TUBB4	NM_006086
105	202163	The CCR4-NOT transcription complex, subunit 8	CNOT8	NM_004779
					106	202187	Phosphoprotein phosphatase 2 is regulated subunit B α abnormal shape	PPP2R5A	NM_006243
107	202197	Myotube element (Myotubularin) associated protein 3	MTMR3	NM_021090
					108	202219	Solute carrier family 6, the member 8	SLC6A8	NM_005629
109	202231	Dendritic cell albumen	GA17	NM_006360
					110	202233	The ubiquinol-cytochrome c reductase hinge protein	UQCRH	NM_006004
111	202242	Stride film 4 superfamily members 2	TM4SF2	NM_004615
					112	202275	Glucose-6-phosphate dehydrogenase	G6PD	NM_000402
113	202279	Karyomit(e) 14 open reading frame 2	C14ORF2	NM_004894

114	202285	The tumour calcium signal transducer 2 of being correlated with		NM_002353
					115	202286	The tumour calcium signal transducer 2 of being correlated with		NM_002353
116	202287	The tumour calcium signal transducer 2 of being correlated with		NM_002353
					117	202298	The inferior complex body of nadh dehydrogenase 1 α, 1	NDUFA1	NM_004541
118	202310	α 1 (I) chain before the I type is collagenolytic		NM_000088
					119	202311	α 1 (I) chain before the I type is collagenolytic		NM_000088
120	202312	Collagen, the I type, α 1	COL1A1	NM_000088
					121	202324	The resident Protein G CP60 of golgi body	GCP60	NM_022735
122	202325	The ATP synthetic enzyme, H+ transhipment, cytopigment F0 complex body, subunit F6	ATP5J	NM_001685
					123	202349	Dystonia 1 is reversed	DYT1	NM_000113
124	202411	Interferon, rabbit, α inducible protein 27	IFI27	NM_005532
					125	202423	Zinc finger protein 22 0	ZNF220	NM_006766
126	202432	Phosphoprotein phosphatase 3 catalytic subunits, the β abnormal shape	PPP3CB	NM_021132
					127	202442	Joint associated protein complex body 3, ∑ 1sub	AP3S1	NM_001284
128	202458	Proteolytic enzyme, Serine, 23	SPUVE	NM_007173
					129	202468	Catenin α-sample 1	CTNNAL1	NM_003798
130	202478	GS3955 albumen	GS3955	NM_021643
					131	202481	Short-chain dehydrogenase reductase enzyme 1	SDR1	NM_004753
132	202503	KIAA0101	KIAA0101	NM_014736
					133	202544	Glia maturation factor, β	GMFB	NM_004124
134	202565	Super villin (supervillin), transcript variant 1	SVIL	NM_003174
					135	202582	The RAN bindin 9	RANBPM	NM_005493
136	202591	Single-stranded DNA binding protein	SSBP	NM_003143
					137	202622	Spinocebellar ataxia (spinocerebellar ataxia) 2	SCA2	NM_002973
138	202642	The structural domain associated protein is transcribed in conversion	TRRAP	NM_003496
					139	202649	Ribosomal protein S19	RPS19	NM_001022
140	202673	Dolichyl-phosphate mannosyltransferase polypeptide 1, catalytic subunit	DPM1	NM_003859
					141	202692	The upstream is in conjunction with transcription factor, rna plymerase i	UBTF	NM_014233

142	202712	Creatine kinase, mitochondrial (mitochondrial) 1	CKMT1	NM_020990
					143	202723	Jaw frame albumen (forkhead box) O1A	FOXO1A	NM_002015
144	202724	Jaw frame albumen O1A	FOXO1A	NM_002015
					145	202735	Emopamil is conjugated protein	EBP	NM_006579
146	202754	KIAA0029 albumen	KIAA0029	NM_015361
					147	202759	A kinases (PRKA) anchorin 2	AKAP2	NM_007203
148	202760	A kinases (PRKA) anchorin 2	AKAP2	NM_007203
					149	202761	Cynapse nuclear expression gene 2	KIAA1011	NM_015180
150	202789	Phospholipase C, γ 1		NM_002660
					151	202820	Aromatic hydrocarbon receptor	AHR	NM_001621
152	202824	Transcriptional elongation factor B (SIII), polypeptide 1	TCEB1	NM_005648
					153	202834	Serine (or halfcystine) proteinase inhibitor, clade A member 8	SERPINA8	NM_000029
154	202841	7-60 albumen	7-60	NM_007346
					155	202848	G protein coupled receptor albumen 6		NM_002082
156	202854	Hypoxanthine phosphoribosyltransferase 1	HPRT1	NM_000194
					157	202860	The KIAA0476 gene product	KIAA0476	NM_014856.
158	202889	Microtubule bindin 7		NM_003980
					159	202890	Microtubule bindin 7		NM_003980
160	202947	Glycophorin C is transcribed variant 1	GYPC	NM_002101
					161	202949	Four and half (four and a half) LIM structural domain 2	FHL2	NM_001450
162	202957	The Lyn substrate 1 that hematopoietic cell is special	HCLS1	NM_005335
					163	203044	KIAA0990 albumen	KIAA0990	NM_014918
164	203053	The sequence 2 of mammary cancer amplification	BCAS2	NM_005872
					165	203133	Albumen displacement complex body β	SEC61B	NM_006808
166	203138	Histone acetyltransferase 1	HAT1	NM_003642
					167	203139	Dead related protein kinase 1	DAPK1	NM_004938
168	203140	B cell CLL lymphoma 6	BCL6	NM_001706
					169	203142	Joint associated protein complex body 3, β 1 subunit	AP3B1	NM_003664
170	203211	KIAA1073 albumen	KIAA1073	NM_016156
					171	203213	Cell division cycle 2, G1 to S and G2 to M		NM_001786

172	203255	Vitiligo associated protein VIT-1	VIT1	NM_018693
					173	203262	The sequence that chromosome x (single) 9928 is expressed	DXS9928E	NM_004699
174	203287	ladinin?1	LAD1	NM_005558
					175	203316	Small nuclear ribonucleoprotein polypeptide E	SNRPE	NM_003094
176	203332	Inositol polyphosphate-5-Phosphoric acid esterase, 145kD	INPP5D	NM_005541
					177	203362	MAD2 (defective, yeast, homologue are stagnated in mitotic division)-sample 1	MAD2L1	NM_002358
178	203371	Nadh dehydrogenase 1 β complex body, 3	NDUFB3	NM_002491
					179	203385	Diacylglycerol kinase, α	DGKA	NM_001345
180	203396	Proteasome (prosome, macropain) subunit, α type, 4	PSMA4	NM_002789
					181	203437	The receptor protein of inferring	PMI	NM_003876
182	203460	Presenilin 1, transcript I-463	PSEN1	NM_007318
					183	203484	Sec61γ	SEC61G	NM_014302
184	203514	Mitogen activated protein kinase kinase kinase 3		NM_002401
					185	203528	The Sema structural domain, Ig structural domain, TM structural domain and short cytoplasmic structure territory, 4D	SEMA4D	NM_006378
186	203531	cullin?5		NM_003478
					187	203581	RAB4, member RAS oncogene family		NM_004578
188	203610	Ring finger protein 15		NM_006355
					189	203613	The inferior complex body of nadh dehydrogenase 1 β, 6	NDUFB6	NM_002493
190	203621	The inferior complex body of nadh dehydrogenase 1 β, 5	NDUFB5	NM_002492
					191	203666	The mesenchymal cell derived factor-1	SDF1	NM_000609
192	203667	Tubulin specificity chaperone a	TBCA	NM_004607
					193	203674	The KIAA0054 gene product; Helicase	KIAA0054	NM_014877
194	203743	Thymus pyrimidine-DNA transglucosylase	TDG	NM_003211
					195	203748	The RNA binding motif, strand interaction protein 1, transcript variant MSSP-2	RBMS1	NM_016839
196	203761	Src sample joint	SLA	NM_006748
					197	203827	Putative protein FLJ10055	FLJ10055	NM_017983
198	203845	The p300CBP correlation factor		NM_003884

199	203882	The transcription factor 3 that Interferon, rabbit stimulates, γ	ISGF3G	NM_006084
					200	203886	fibulin?2	FBLN2	NM_001998
201	203893	Suprarenal gland albumin A D-004	LOC51578	NM_016283
					202	203922	Cytochrome b-245, beta polypeptides		NM_000397
203	203936	Matrix metalloproteinase 9	MMP9	NM_004994
					204	203940	KIAA1036 albumen	KIAA1036	NM_014909
205	203960	HSPCO34 albumen	LOC51668	NM_016126
					206	203965	Ubiquitin-specific protease 20	USP20	NM_006676
207	204014	Dual specificity Phosphoric acid esterase 4	DUSP4	NM_001394
					208	204015	Dual specificity Phosphoric acid esterase 4	DUSP4	NM_001394
209	204049	The KIAA0680 gene product	KIAA0680	NM_014721
					210	204067	Sulfite oxidase		NM_000456
211	204082	Pre B cell leukemia transcription factor 3	PBX3	NM_006195
					212	204141	Tubulin, beta polypeptides	TUBB	NM_001069
213	204145	FSHD district gene 1	FRG1	NM_004477
					214	204146	The RAD51 interaction protein		NM_006479
215	204158	The T cell, immunomodifier 1	TCIRG1	NM_006019
					216	204168	Microsome glutathione S-transferase 2	MGST2	NM_002413
217	204170	CDC28 protein kinase 2	CKS2	NM_001827
					218	204215	Putative protein MGC4175		NM_024315
219	204294	Aminomethyltransferase	AMT	NM_000481
					220	204351	S100 calcium binding protein P	S100P	NM_005980
221	204361	The SKAP55 homologue	SKAP-Hom	NM_003930
					222	204362	The SKAP55 homologue	SKAP-Hom	NM_003930
223	204411	KIAA0449 albumen	KIAA0449	NM_017596
					224	204416	ApoC-I	APOC1	NM_001645
225	204528	Nucleosome assembly protein 1-sample 1	NAP1L1	NM_004537
					226	204640	Spotted type POZ albumen	SPOP	NM_003563
227	204642	The endothelium differentiation, schwann's sheath ester G-albumen-coupled receptor, 1	EDG1	NM_001400
					228	204652	Nuclear is breathed the factor 1	NRF1	NM_005011
229	204694	Alpha-fetoprotein	AFP	NM_001134

230	204698	The gene that Interferon, rabbit stimulates	ISG20	NM_002201
					231	204729	Syntaxin 1A	STX1A	NM_004603
232	204766	Nudix type motif 1	NUDT1	NM_002452
					233	204767	Flap structure specific nucleic acid restriction endonuclease 1		NM_004111
234	204777	T cytodifferentiation albumen is transcribed variant a	MAL	NM_002371
					235	204863	The interleukin 6 signal transducer		NM_002184
236	204864	The interleukin 6 signal transducer	IL6ST	NM_002184
					237	204881	UDP-glucose Ceramide glucosyltransferase	UGCG	NM_003358
238	204885	Mesothelin transcribes variant 1	MSLN	NM_005823
					239	204905	Eukaryotic translation EF-1 ε 1	EEF1E1	NM_004280
240	204923	Chromosome x q25-26.1 goes up 753P9		NM_018990
					241	204950	KIAA0955 albumen	KIAA0955	NM_014959
242	204951	Ras homologous gene family, member H	ARHH	NM_004310
					243	204955	Contain sushi multiple albumen, X chromosome	SRPX	NM_006307
244	204966	The angiogenesis inhibitor 2 that brain is special	BAI2	NM_001703
					245	204989	Integrate plain (integrin) β 4	ITGB4	NM_000213
246	204990	Integrate plain β 4	ITGB4	NM_000213
					247	205033	Defensin (defensin), α 1, the marrow correlated series	DEFA1	NM_004084
248	205087	DKFZP566K023 albumen		NM_015485
					249	205108	Apolipoprotein B	APOB	NM_000384
250	205133	Heat-shocked 10kD albumen 1	HSPE1	NM_002157
					251	205176	It is conjugated protein to integrate plain β 3	ITGB3BP	NM_014288
252	205213	The KIAA0050 gene product	ACAP1	NM_014716
					253	205270	Lymphocyte cytoplasmic protein 2	LCP2	NM_005565
254	205323	The transcription factor 1 that metal is regulated	MTF1	NM_005955
					255	205335	Signal recognition particle 19kD	SRP19	NM_003135
256	205336	Parvalbumin	PVALB	NM_002854
					257	205339	TAL1 (SCL) interrupted gene seat	SIL	NM_003035
258	205361	prefoldin?4		U41816
					259	205403	Interleukin 1 receptor, the II type	IL1R2	NM_004633
260	205453	With source capsule B2	HOXB2	NM_002145

261	205467	caspase?10	CASP10	NM_001230
					262	205600	With source capsule B5	HOXB5	NM_002147
263	205601	With source capsule B5	HOXB5	NM_002147
					264	205608	Angiogenin 1		NM_001146
265	205609	Angiogenin 1	ANGPT1	NM_001146
					266	205644	Small nuclear ribonucleoprotein polypeptide G	SNRPG	NM_003096
267	205671	MHC, II class DO β	HLA-DOB	NM_002120
					268	205807	tuftelin?1	TUFT1	NM_020127
269	205849	Gamma amino butyric acid A acceptor β 3 transcribes variant 1	GABRB3	NM_006294
					270	205967	The H4 histone family, member G	H4FG	NM_003542
271	206066	RAD51 (S.cerevisiae) homologue C	RAD51C	NM_002876
					272	206148	The interleukin-13 acceptor, α	IL3RA	NM_002183
273	206150	Tumor necrosis factor receptor super family, the member 7	TNFRSF7	NM_001242
					274	206177	Arginase, liver	ARG1	NM_000045
275	206219	Vav 1 oncogene	VAV1	NM_005428
					276	206245	NS1 is conjugated protein	NS1-BP	NM_006469
277	206289	With source capsule A4	HOXA4	NM_002141
					278	206298	Putative protein is from s 23549 and 23762	LOC58504	NM_021226
279	206618	Interleukin 18 acceptor 1	IL18R1	NM_003855
					280	206637	KIAA0001	KIAA0001	NM_014879
281	206674	The Fms Tyrosylprotein kinase 3 of being correlated with	FLT3	NM_004119
					282	206723	G protein coupled receptor Edg-4		AF233092
283	206790	The inferior complex body of nadh dehydrogenase 1 β, 1	NDUFB1	NM_004545
					284	206868	The KIAA0189 gene product	KIAA0189	NM_014725
285	206874	The serine threonine kinases that Ste20 is relevant		NM_014720
					286	206958	UPF3	UPF3	NM_023011
287	207072	Interleukin 18 acceptor accessory protein	IL18RAP	NM_003853
					288	207111	Contain Egf sample assembly, Saliva Orthana sample, hormone receptor sample sequence 1	EMR1	NM_001974
289	207127	Heterogeneous nuclear ribonucleoprotein H3	HNRPH3	NM_021644
					290	207163	V-akt mouse thymoma virus oncogene homologue 1	AKT1	NM_005163

291	207165	Hyaluronan mediated motility receptor is transcribed variant 2	HMMR	NM_012485
					292	207266	The RNA binding motif, strand interaction protein 1 is transcribed variant MSSP-3	RBMS1	NM_016837
293	207287	Putative protein FLJ14107	FLJ14107	NM_025026
					294	207335	1 member 7 of solute belongings family	SLC1A7	NM_006671
295	207540	Spleen tyrosine kinase	SYK	NM_003177
					296	207551	Male specific fatal-3 (fruit bat)-samples 1	MSL3L1	NM_006800
297	207568	Cholinergic receptor, nicotine, α polypeptide 6	CHRNA6	NM_004198
					298	207571	Basilar membrane inductive gene	ICB-1	NM_004848
299	207573	The ATP synthetic enzyme, H+ transhipment, plastosome F1F0, subunit g	ATP5JG	NM_006476
					300	207777	Nucleosome Protein S p140	SP140	NM_007237
301	207826	DNA binding inhibitors 3, dominance negative interaction spiral-ring-coilin	ID3	NM_002167
					302	207845	Later stage promotes complex body 10	APC10	NM_014885
303	207935	Keratin sulfate 13	KRT13	NM_002274
					304	207974	S phase kinase-associated protein 1A (p19A)	SKP1A	NM_006930
305	208091	Putative protein DKFZp564K0822		NM_030796
					306	208130	The thromboxane A synthetase 1	TBXAS1	NM_030984
307	208270	The arginyl aminopeptidase	RNPEP	NM_020216
					308	208310	Follistatin sample 1	FSTL1	NM_007085
309	208325	Lymphocyte sudden turn of events oncogene	LBC	NM_006738
					310	208414	With source capsule B3	HOXB3	NM_002146
311	208420	Repressor 6 homologues of Ty (S.cerevisiae)	SUPT6H	NM_003170
					312	208549	Prothymosin a14	LOC51685	NM_016171
313	208598	Upstream controlling element conjugated protein 1	UREB1	NM_005703
					314	208621	Villin 2	ezrin	NM_003379
315	208622	Villin 2	ezrin	NM_003379
					316	208623	Cytovillin 2	VIL2	NM_003379
317	208629	Hydroxyl acyl group-coa dehydrogenase 3-keto acyl base-coenzyme A thiolaseenoyl-CoA hydratase α subunit		U04627

318	208633	Actin binding protein; Macrophin		AB029290
					319	208643	Ku autoimmunization antigen gene		J04977
320	208656	Cyclin I	CYC1	AF135162
					321	208667	The tumor suppressor gene ST13 that infers	ST13	U17714
322	208672	Splicing factor is rich in arginine Serine 3		BC000914
					323	208679	PNAS-139		AF279893
324	208695	Ribosomal protein L 39	RPL39	BC001019
					325	208745	The ATP synthetic enzyme, H+ transhipment, plastosome F1F0, g subunit		AA917672
326	208746	F1F0 type ATP synthetic enzyme subunit g		AF070655
					327	208754	DKFZp762G106		AL162068
328	208808	The albumen 2 of high mobility group		BC000903
					329	208819	Mel transforms oncogene-RAB8 homologue		BC002977
330	208831	KIAA0162		D79984
					331	208894	MHC II type HLA-DR-α		M60334
332	208900	Topoisomerase I		W025108
					333	208904	Ribosomal protein S28	RPS28	BC000354
334	208905	Cytochrome c		BC005299
					335	208919	Be similar to putative protein FLJ13052		BC001709
336	208927	Spotted type POZ albumen		BC001269
					337	208956	The deoxyguanosine triphosphate enzyme	DUT	U62891
338	208966	Interferon, rabbit, but γ inducible protein 16		AF208043
					339	208975	Importin β subunit		L38951
340	208977	Tubulin, β, 2		BC004188
					341	208981	PECAM		NM_000442
342	209022	Similar to the autonomously replicating sequence height	ARS	AK026678
					343	209063	Polyadenylic acid is conjugated protein-interaction protein 1		BF248165
344	209066	The plastosome ubiquinone is conjugated protein		M26700
					345	209089	RAB5A, member RAS oncogene family		BC001267
346	209119	Nuclear receptor subtribe 2, the F group, the member 2		AV703465
					347	209120	Nuclear receptor subtribe 2, the F group, the member 2		AV703465

348	209122	Fat differentiation associated protein		NM_001122
					349	209138	Immunoglobulin (Ig) λ locus		FLM87790
350	209172	Centromere protein F		U30872
					351	209191	Be similar to tubulin, β, 4		BC002654
352	209209	The mitogen induced gene	mig-2	AW469573
					353	209210	The mitogen induced gene	mig-2	Z24725
354	209227	The prostate cancer tumor suppressor gene of inferring		NM_006765
					355	209228	The prostate cancer tumor suppressor gene of inferring		NM_006765
356	209239	The nf 1 of κ chain polypeptide gene enhanser in the B cell	p105	M55643
					357	209257	Chondroitin sulfate proteoglycan 6	bamacan	NM_005445
358	209258	Chondroitin sulfate proteoglycan 6	bamacan	NM_005445
					359	209270	Ln S B3 chain	LAMB3	NM_000228
360	209282	Protein kinase D2		NM_016457
					361	209289	Nf IB		NM_005596
362	209295	TRAIL acceptor 2		AF016266
					363	209303	Nadh dehydrogenase Fe-S albumen 4		NM_002495
364	209309	Zinc-α 2-glycoprotein		D90427
					365	209324	The retina of G-protein signal transduction enriches conditioning agent 16	hRGS-r	NM_002928
366	209325	The retina of G-protein signal transduction enriches conditioning agent	hRGS-r	NM_002928
					367	209362	RNA polymerase B inhibitor, the yeast homologue	SRB7	NM_004264
368	209369	1,2-ring-Inositol Monophosphate phosphodiesterase	ANX3	NM_005139
					369	209372	Tubulin, beta polypeptides		BC001352
370	209377	Thyroid Hormone Receptors interaction protein 7		AF274949
					371	209385	The proline(Pro) synthetic enzyme of corotation record		AL136616
372	209411	The conjugated protein GGA3 of ADP ribosylation factor		AF219139
					373	209443	Proteinase inhibitor, clade A member 5		NM_000624
374	209471	Farnesyl-protein transferase, the α subunit		NM_002027
					375	209492	The ATP synthetic enzyme, H+ transhipment, plastosome F0 complex body, subunit e		BC003679
376	209500	Tumor necrosis factor superfamily, the member 13	TNSF13	NM_003808
					377	209507	Replication protein A 3		NM_002947

378	209520	Nuclear cap binding protein subunit 1		NM_002486
					379	209560	δ sample homologue		NM_003836
380	209585	Multiple inositol polyphosphate Phosphoric acid esterase		NM_004897
					381	209628	Putative protein P15-2		NM_018698
382	209662	Centrin, EF hand albumen, 3		NM_004365
					383	209679	Putative protein from 643		NM_020467
384	209687	The mesenchymal cell derived factor-1		U19495
					385	209730	The Sema structural domain, Ig, short basic domain, excretory, 3F		U38276
386	209734	Hematopoietic proteins 1		NM_005337
					387	209810	Curosurf associated protein B	SP-B	NM_000542
388	209827	Interleukin-11 6	IL16	NM_004513
					389	209838	Thryoid receptor interaction protein 15		AF212227
390	209868	The RNA binding motif, strand interaction protein 1	SCR2	NM_002897
					391	209907	Intersectin 2 is long special-shaped	ITSN2	AF182198
392	210093	The mago-nashi homologue, propagation is correlated with		NM_002370
					393	210097	But vitamin A acid arrestin		AF130102
394	210137	The dCMP desaminase		BC001286
					395	210139	Sphingophospholipid 4 protein 22 on every side	GAS3	L03203
396	210180	Splicing factor is rich in arginine Serine 10		U87836
					397	210233	The interleukin 1 receptor accessory protein	IL1RAP	AF167343
398	210235	LAR interaction protein 1a		U22815
					399	210283	Be similar to polyadenylic acid binding protein interactions 1		BC005295
400	210284	TAK1 conjugated protein 2		AF241230
					401	210347	C2H2 type zinc finger protein		AF080216
402	210448	Ionic ATP acceptor P2X5b		U49396
					403	210453	DKFZp566G013		AL050277
404	210510	Solvable neuropilin-1		AF145712
					405	210563	FLICE sample consistence albumen short-form		U97075
406	210598			AF130051
					407	210615	Neuropilin-1 solvable special-shaped 11	NRP1	AF280547

408	210633	Acid Keratin sulfate-10	KRT10	M19156
					409	210639	Apoptosis-related protein	APG5L	AF293841
410	210649	BRG1 binding factor 250a	BAF250a	AF231056
					411	210666	Iduronic acid 2-sulfatase	IDS	NM_000202
412	210785	The basilar membrane induced gene	ICB-1beta	AB035482
					413	210786	Fu Luode (Friend) leukosis virus integral protein 1	FLI-1	M93255
414	210840	IQ motif cont ' g GTP enzyme activation albumen 1		D29640
					415	210854	The GABA norepinephrine transporter		U17986
416	210859	CLN3 albumen	CLN3	AF077973
					417	210982	MHC II type HLA-DRA	HLA-DRA	M60333
418	211000	Gp130 carries the soluble form of rheumatoid arthritis antigen peptide	gp130-RAPS	AB015706
					419	211133	White corpuscle Ig sample acceptor subtribe B member 3	LILRB3	NM_00864
420	211430	Anti-hepatitis A IgG		M87789
					421	211445	FKSG17	FKSG17	AF315951
422	211487	Ribosomal protein S17		BC004886
					423	211535	Fibroblast growth factor receptor	FGFR	M60485
424	211645	Immunoglobulin kappa chain VK-1	IgK	M85256
					425	211730	Polysaccharase (RNA) II (the DNA guiding) polypeptide L		BC005903
426	211743	Proteoglycan 2, marrow		BC005929
					427	211747	The U6snRNA Sm sample albumen of being correlated with		BC005938
428	211762	Nuclear translocation protein alpha 2		BC005978
					429	211858	The special-shaped L2 of guanine nucleotide binding protein Gs α subunit		AF088184
430	211915	The β tubulin	TUB4q	U83110
					431	211921	The fetal thymus prothymosin		AF348514
432	211932	Heterogeneous albumen is similar to rat spiral unstability (helix destabilizing) albumen		BE867771
					433	211938	Putative protein PRO1843		BF247371
434	211976	FLJ21862fis		AK026168
					435	211985	Matrix Gla albumen		AI653730
436	212007	Contain UBX structural domain-1		D87684

437	212012	KIAA0230		AF200348
					438	212013	KIAA0230		AF200348
439	212027	S164 albumen		BE466128
					440	212181	IP2 polyphosphate phosphohydrolase 2	NUDT4	AF191654
441	212201	KIAA0692 albumen		AB014592
					442	212248	FLJ20738fis		AI972475
443	212269	Minichromosome keeps defective 3-associated protein	GANP	AJ010089
					444	212277	KIAA0647 albumen		AB014547
445	212283	est：tg49b04.x1		AF016903
					446	212285	est：tg49b04.x1		AF016903
447	212287	KIAA0160 albumen		BF382924
					448	212298	neuropilin?1		NM_003873
449	212311	KIAA0746 albumen		AB018289
					450	212382	FLJ11918fis		AK021980
451	212384	HLA-B associated retroviral basis-1		BG341380
					452	212385	FLJ11918fis		AK021980
453	212386	FLJ11918fis		AK021980
					454	212387	FLJ11918fis		AK021980
455	212426	Tyrosine 3-monooxygenase Tryptophan 5-monooxygenase activated protein, the θ polypeptide		BF033313
					456	212451	KIAA0256		D87445
457	212531	lipocalin?2	LCN2	NM_005564
					458	212549	DKFZp586N1323		AI149535
459	212561	KIAA1091		AA349595
					460	212570	KIAA0830		AL573201
461	212571	KIAA1564		U00955
					462	212573	KIAA0830		AL573201
463	212578	Ribosomal protein S17		BF026595
					464	212583	The KIAA0560 gene product		AB011132
465	212630	The sec6 homologue		AF055006
					466	212664	Tubulin, β, 5		NM_006087

467	212740	Phosphoinositide-3-kinases, modulability sub 4	p150	BF740111
					468	212757	FLJ22656fis		BF111268
469	212771	DKFZp564A026		AU150943
					470	212833	The TB1 gene		M74089
471	212860	DKFZp667O2416		BG168720
					472	212878	Kinesin 2		AA284075
473	212907	hbc647		AI972416
					474	212927	KIAA0594		AB011166
475	212964	DKFZp434D1023		AB028943
					476	213020	DKFZp566B213	GOSR1	NM_004871
477	213028	With the conjugated protein relevant nf of κ B		AI887378
					478	213047	SET be shifted (translocation)		AI278616
479	213061	KIAA0251		AA643304
					480	213062	KIAA0251		AA643304
481	213074	DKFZp564D156		BG545769
					482	213129	Glycine diced system albumen H		BE908931
483	213134	BTG family, the member 3	BTG3	NM_-006806
					484	213147	Homology frame A10		NM_018951
485	213150	Homology frame A10		NM_018951
					486	213159	KIAA0805		AB018348
487	213178		MAPK8IP3	NM_015133
					488	213188	Be similar to T15138hyp albumen T28F2.4 slightly		NM_032778
489	213231	Dystrophia myotonica-contain WD to repeat motif		L19267
					490	213253	The structure smaint of karyomit(e) 2, yeast-sample 1	SMC2L1	AU154486
491	213287	The encoding gene of acid (I type) cytokeratin 10		X14487
					492	213311	KIAA1049 albumen		BF000251
493	213338	DKFZP586E1621		BF062629
					494	213414	Ribosomal protein S19		BE259729
495	213423	The prostate cancer tumor inhibitor of inferring		AI884858
					496	213479	neuronal?pentraxin?II	NPTX2	NM_002523
497	213502	Immunoglobulin (Ig) λ-sample polypeptide 3		X03529

498	213539	CD3D antigen, δ polypeptide (TiT3 complex body)	CD3D	NM_000732
					499	213553	ApoC-I		W79394
500	213567	23728 sequences		BF431965
					501	213599	Opa-interaction protein 5		BE045993
502	213687	The hypothalamus albumen HSMNP1 of Biao Zhenging not		BE968801
					503	213700	Putative protein FLJ10803		AA554945
504	213727	Putative protein FLJ11585		AI743654
					505	213754	Polyadenylic acid is conjugated protein-interaction protein 1		AW613203
506	213811	Transcription factor 3		BG393795
					507	213843	Attached protein B AP31BAP29		AW276522
508	213867	The β Actin muscle		AA809056
					509	213911	The H2A histone family, member Z		BF718636
510	213941	Ribosomal protein S7		AI970731
					511	214003	Ribosomal protein S20		BF184532
512	214051	Extrasin beta		BF677486
					513	214097	Ribosomal protein S21		AW024383
514	214124	DKFZp434B2027		AL043487
					515	214129	KIAA0477		AI821791
516	214143	Ribosomal protein L 24		AI560573
					517	214224	Albumen NIMA-interacts, and 4	parvulin	BE674061
518	214264	est：tt46h03		AI656610
					519	214288	Proteasome (prosome, macropain) subunit, β type, 1		W86293
520	214292	Integrate plain β 4		AA808063
					521	214298	septin?2		AL568374
522	214352	V-Ki-ras2Kirsten rat sarcoma 2 viral oncogene homologues		BF673699
					523	214377	est：UI-H-BI4-aop-e-12-0-UI.s1		BF508685
524	214433	Selenium conjugated protein 1	SELENBP1	NM_003944
					525	214494	The cell matrix cell adhesion modulating agent	CMAR	NM_005200
526	214499	Bcl-2-associated transcription factor short-form		AF249273

527	214617	Perforin 1		AI445650
					528	214661	On 4p16.3,, has homology with hyp S. pombe gene near HD		R06783
529	214696	24659 sequences		AF070569
					530	214721	DKFZp762L106		AL162074
531	214800	Basal transcription factor 3		R83000
					532	214820	The N143 of transcription unit		AJ002572
533	214905	EUROIMAGE46866		AL109674
					534	215038	The Huntingtin interaction protein		AF049103
535	215073	Nuclear receptor family 2, the F hypotype, the member 2		AL554245
					536	215096	Esterase Dformylglutathione lytic enzyme		AU145746
537	215121	Immunoglobulin (Ig) λ locus		AA680302
					538	215127	The RNA binding motif, strand interaction prot1		AL517946
539	215136	DKFZp564C0482		AL050353
					540	215147	23712 sequences		AF007147
541	215171	NT2RP3000341		AK023063
					542	215227	Acid phosphatase 1, solvable		BG035989
543	215379	Immunoglobulin (Ig) λ connects 3		AV698647
					544	215380	FLJ11717fis		AK021779
545	215399	Amplification in osteosarcoma (osteosarcoma)		AI683900
					546	215446	Lysyloxidase	LOX	L16895
547	215493	3 ends of the BTN2A1 gene of coding butyrophilin 2A		AL121936
					548	215691	HSPCO34 albumen		AV702994
549	215764	Joint associated protein complex body 2, α 2 subunits		AA877641
					550	215812	The creatine translocator	SLC6A10	U41163
551	215905	The 40kDa albumen that U5snRNP-is special		AL157420
					552	216207	The variable 1-13 of immunoglobulin (Ig) κ		AW408194
553	216348	Transcription factor AP-1-2		AL049693
					554	216384	Prothymosin	PTMA	AF257099
555	216641	ladinin	LAD	U58994

556	216833	Courageous and upright glycoprotein (glycophorin) HeP2		U05255
					557	216899	PAC RP5-1139P1 is from 7p15-p21		AC003999
558	216977	The special A albumen of U2snRNP-, alternative transcription basis 3		AJ130972
					559	217013	PAC RP4-604G5 is from 7q22-q31.1		AC004522
560	217014	PAC RP4-604G5 is from 7q22-q31.1		AC004522
					561	217022	Ig A1-A2 λ hybrid GAU heavy chain		S55735
562	217106	The dimethyladenosine transferring enzyme of inferring		AF091078
					563	217122	Stromatin enzyme female reproductive tract (Rep tract) MIFR12	MMP2122A	AL031282
564	217249	BAC CTB-162B4 is from 4		AC004544
					565	217293	Angiogenin 1		AF209975
566	217329	Cytochrome c oxidase subunit VIIb pseudogene	COX7BP1	AF042164
					567	217336	RP5-858M22 on the karyomit(e) 20		AL118510
568	217410	FLJ11524fis		AK021586
					569	217419	FLJ11524fis		AK021586
570	217491	Cytochrome c oxidase subunit VIIc pseudogene	COX7CP1	AF042165
					571	217749	Coat protein γ-cop	LOC51137	NM_016128
572	217750	Putative protein FLJ13855		NM_023079
					573	217754	The nucleolar RNA helicase of inferring	NOH61	NM_019082
574	217769	Putative protein	HSPC014	NM_015932
					575	217773	The inferior complex body of nadh dehydrogenase 1 α, 4	NDUFA4	NM_002489
576	217801	The ATP synthetic enzyme, H+ transhipment, plastosome F1 mixture, epsilon subunit	ATP5E	NM_006886
					577	217812	High glucose regulated protein 8	HGRG8	NM_016258
578	217825	CGI-76 albumen		AF151039
					579	217833	The NS1-associated protein 1		NM_006372
580	217838	RNB6	RNB6	NM_016337
					581	217843	HSPC126 albumen	HSPC126	NM_014166
582	217853	Putative protein FLJ13732 is similar to tensin	TENC1	NM_022748
					583	217860	The inferior complex body of nadh dehydrogenase 1 α, 10	NDUFA10	NM_004544
584	217866	Putative protein FLJ12529	FLJ12529	NM_024811

585	217875	Stride film, prostate gland male sex hormone inductive RNA	TMEPAI	NM_020182
					586	217877	Putative protein SP192	SP192	NM_021639
587	218003	FK506-conjugated protein 3	FKBP3	NM_002013
					588	218039	HQ0310PRO0310p1	LOC51203	NM_016359
589	218058	CpG is conjugated protein	CGBP	NM_014593
					590	218062	Cdc42 effect protein 4; The binding substances of Rho GTPases 4	CEP4	NM_012121
591	218103	Putative protein FLJ20062	FLJ20062	NM_017647
					592	218116	Stem cell knurl related antigen 59	LOC51759	NM_016520
593	218117	Ring-box 1	RBX1	NM_014248
					594	218175	Putative protein FLJ22471	FLJ22471	NM_025140
595	218188	Mitochondrial inner membrane translocase 13 (yeast) homologue B	TIMM13B	NM_012458
					596	218213	Karyomit(e) 11 open reading frame 10	C11orf10	NM_014206
597	218241	Gorky's autoantigen, golgin subfamily a, 5	GOLGA5	NM_005113
					598	218256	Nucleoporin p54	NUP54	NM_017426
599	218259	KIAA1243 albumen	KIAA1243	NM_014048
					600	218274	Putative protein FLJ10415	FLJ10415	NM_018089
601	218276	Contain the WW domain gene	WW45	NM_021818
					602	218280	The H2A histone family, member O	H2AFO	NM_003516
603	218283	Kiaa-iso albumen	LOC51188	NM_016305
					604	218288	Putative protein MDS025	MDS025	NM_021825
605	218334	Putative protein FLJ23445	FLJ23445	NM_025075
					606	218339	HSPC158 albumen	HSPC158	NM_014180
607	218350	geminin	LOC51053	NM_015895
					608	218367	Ubiquitin-specific protease 21	USP21	NM_012475
609	218368	I type transmembrane protein Fn14	FN14	NM_016639
					610	218373	Hyp albumen FLJ13258 is similar to the toes of fusion	FLJ13258	NM_022476
611	218395	Putative protein FLJ13433	FLJ13433	NM_022496
					612	218397	Putative protein FLJ10335	FLJ10335	NM_018062
613	218447	DC13 albumen	DC13	NM_020188
					614	218467	X 003 albumen	MDS003	NM_020232

615	218482	DC6 albumen	DC6	NM_020189
					616	218543	Putative protein FLJ22693	FLJ22693	NM_022750
617	218563	The inferior complex body of nadh dehydrogenase 1 α, 3	NDUFA3	NM_004542
					618	218576	Two-fold specificity phosphotase 12	DUSP12	NM_007240
619	218603	Fruit bat headcase homologue	hHDC	NM_016217
					620	218605	Putative protein FLJ23182	FLJ23182	NM_022366
621	218643	Postsynaptic PROTEIN C RIPT	CRIPT	NM_014171
					622	218660	Dysferlin, limb girdle type muscular dystrophy 2B	DYSF	NM_003494
623	218671	The atpase inhibitor precursor	LOC51189	NM_016311
					624	218801	UDP-glucose: glucoprotein glucose base transferring enzyme 2	FLJ10873	NM_020121
625	218830	Ribosomal protein L 26 homologues	LOC51121	NM_016093
					626	218873	Putative protein FLJ20203	FLJ20203	NM_017710
627	218936	HSPC128 albumen	HSPC128	NM_014167
					628	218937	Putative protein FLJ20417	FLJ20417	NM_017810
629	218946	HIRIP5 albumen	HIRIP5	NM_015700
					630	219008	Putative protein FLJ21820	FLJ21820	NM_021925
631	219030	CGI-121 albumen	LOC51002	NM_016058
					632	219032	Opsin 3 (encephalopsin)	OPN3	NM_014322
633	219056	Putative protein FLJ11712	FLJ11712	NM_024570
					634	219105	Origin recognition complex, subunit 6-sample	ORC6L	NM_014321
635	219110	GAR1 albumen	GAR1	NM_018983
					636	219163	Putative protein FLJ20079	FLJ20079	NM_017656
637	219218	Putative protein FLJ23058	FLJ23058	NM_024696
					638	219286	Putative protein FLJ12479	FLJ12479	NM_022768
639	219293	Putative protein	PTD004	NM_013341
					640	219347	Putative protein FLJ10956	FLJ10956	NM_018283
641	219452	Infer pepx	LOC64174	NM_022355
					642	219506	Putative protein FLJ23221	FLJ23221	NM_024579
643	219507	Putative protein	LOC51319	NM_016625
					644	219546	Putative protein DKFZp434P0116		NM_017593
645	219759	Aminopeptidase	LOC64167	NM_022350

646	219765	Putative protein FLJ12586	FLJ12586	NM_024620
					647	219816	Putative protein FLJ10482	FLJ10482	NM_018107
648	219819	HSPC007 albumen	HSPC007	NM_014018
					649	219906	Putative protein FLJ10213	FLJ10213	NM_018029
650	220001	The peptidyl arginine deiminase, V-type	PAD	NM_012387
					651	220023	The apolipoprotein B48 acceptor	APOB48R	NM_018690
652	220052	TERF1 (TRF1)-interaction nf 2	TINF2	NM_012461
					653	220060	Putative protein FLJ20641	FLJ20641	NM_017915
654	220155	Putative protein FLJ13441	FLJ13441	NM_023924
					655	220199	Putative protein FLJ12806	FLJ12806	NM_022831
656	220386	Karyomit(e) 2 open reading frame 2	C2ORF2	NM_019063
					657	220404	PRO0611 albumen	PRO0611	NM_014076
658	220416	Putative protein FLJ21472	FLJ21472	NM_024837
					659	220558	Full hematopoiesis is to express	PHEMX	NM_005705
660	220671	The carbon degraded product checks the 4-sample	CCRN4L	NM_012118
					661	220741	Inorganic pyrophosphatase	SID6-306	NM_006903
662	220755	G8 albumen	G8	NM_016947
					663	220864	CGI-39 albumen; Protein G RIM19 is regulated in necrocytosis	LOC51079	NM_015965
664	220942	Putative protein, the estradiol inductive	E2IG5	NM_014367
					665	221009	PPAR (γ) angiogenin associated protein	PGAR	NM_016109
666	221143	Replication protein A comp 34kd subunit homologue Rpa4	HSU24186	NM_013347
					667	221253	Putative protein MGC3178	MGC3178	NM_030810
668	221432	Putative protein NPD016	NPD016	NM_031212
					669	221434	Putative protein DC50	DC50	NM_031210
670	221505	Putative protein MGC5350		AW612574
					671	221509	Auto-regulating System of Density of Heavy Medium albumen	SMAP-3	AB014731
672	221528	Be similar to putative protein FLJ11656		BC000143
					673	221577	The prostate gland differentiation factor		AF003934
674	221593	Be similar to ribosomal protein L 31		BC001663
					675	221599	Be similar to PTD015 albumen		BC002752
676	221620	Brain my025		AF061264

The reference tabulation

5,242,974?5,561,071

5,384,261?5,571,639

5,405,783?5,593,839

5,412,087?5,599,695

5,424,186?5,624,711

5,429,807?5,658,734

5,436,327?5,700,637

5,445,934?6,004,755

5,472,672?6,133,305

5,527,681?6,218,114

5,529,756?6,218,122

5,532,128?6,271,002

5,545,531?6,271,210

5,554,501?6,284,764

5,556,752?6,306,897

Adnane?et?al.(2000)″Inhibition?of?farnesyltransferase?increasesTGFbeta?type?II?receptor?expression?and?enhances?the?responsivenessof?human?cancer?cells?to?TGFbeta″Oncogene?19：5525-5533

Albertson?(1984)EMBO?J.3：1227-1234

Alsina?et?al.(2003)″Farnesyltransferase?inhibitor?FTI-R115777is?well?tolerated，induces?stabilization?of?disease，and?inhibitsfarnesylation?and?oncogenic/tumor?survival?pathways?in?patients?withadvanced?multiple?myeloma″Eur.J.Haematol.70：269

Brunner?et?al.(2003)″Farnesyltransferase?inhibitors：anoverview?of?the?results?of?preclinical?and?clinical?investigations″Cancer?Res.63：5656-5668

Bullinger?et?al.(2004)″Use?of?gene-expression?profiling?toidentify?prognostic?subclasses?in?adult?acute?myeloid?leukemia″N.Engl.J.Med.350：1605-1616

Carr?et?al.(1991)″Interaction?of?the?regulatory?subunit?(RII)ofcAMP-dependent?protein?kinase?with?RII-anchoring?proteins?occursthrough?an?amphipathic?helix?binding?motif″J.Biol.Chem.266：14188-14192

Chang?et?al.(2003)″Gene?expression?profiling?for?the?predictionof?therapeutic?response?to?docetaxel?in?patients?with?breast?cancer″Lancet?362：362-369

Cheok?et?al.(2003)″Treatment-specific?changes?in?geneexpression?discriminate?in?vivo?drug?response?in?human?leukemiacells″Nat.Genet.34：85-90

Cortes?et?al.(2003)″Efficacy?of?the?farnesyl?transferaseinhibitor?R115777in?chronic?myeloid?leukemia?and?other?hematologicmalignancies″Blood?101：1692-1697

Cox?et?al.(2002)″Farnesyltransferase?inhibitors：promises?andrealities″Curr.Opin.Pharmacol.2：388-393

Debernardi?et?al.(2003)″Genome-wide?analysis?of?acute?myeloidleukemia?with?normal?karyotype?reveals?a?unique?pattern?ofhomeobox?gene?expression?distinct?from?those?with?translocation-mediated?fusion?events″Genes?Chrom.Cancer?37：149-158

End?et?al.(2001)″Characterization?of?the?antitumor?effects?ofthe?selective?farnesyl?protein?transferase?inhibitor?R115777in?vivoand?in?vitro″Cancer?Res.61：131-137

EP?430,402

Esteva?et?al.(2003)″Phase?III?study?of?weekly?docetaxel?andtrastuzumab?for?patients?with?HER-2-overexpressing?metastaticbreast?cancer″J?Clin?Oncol.20：1800-1808.

Foisner?et?al.(1991)″Protein?kinase?A-and?protein?kinase?C-regulated?interaction?of?plectin?with?lamin?B?and?vimentin″Proc.Natl.Acad.Sci.USA?88：3812-3816

Gene?Therapy?Protocols，edited?by?Paul?D.Robbins，Humanpress，Totowa?NJ，1996

Harousseau?et?al.(2003)″Zarnestra ^TM?(R115777)in?patientswith?relapsed?or?refractory?acute?myelogenous?leukemia?(AML)：results?of?a?multicenter?phase?2study″Blood?102：176a?Abstract?614

Hofmann?et?al.(2002)″Relation?between?resistance?ofPhiladelphia-chromosome-positive?acute?lymphoblastic?leukaemia?tothe?tyrosine?kinase?inhibitor?STI571and?gene-expression?profiles：agene-expression?study″Lancet?359：481-486

Jensen?et?al.(2001)″Estrogen?receptors?and?proliferationmarkers?in?primary?and?recurrent?breast?cancer″Proc?Natl?Acad?SciU?S?A.98：15197-15202.

Jiang?et?al.(2000)″The?phosphoinositide?3-OH?kinase/AKT2pathway?as?a?critical?target?for?farnesyltransferase?inhibitor-inducedapoptosis″Mol.Cell.Biol.20：139-148

Johnston?et?al.(2003)″Phase?II?study?of?the?efficacy?andtolerability?of?two?dosing?regimens?of?the?farnesyl?transferaseinhibitor，R115777，in?advanced?breast?cancer″J.Clin.Oncol。21：2492-2499

Kallioniemi?(1992)Proc.Natl.Acad.Sci.USA?89：5321-5325

Karp?et?al.(2001)″Clinical?and?biologic?activity?of?thefarnesyltransferase?inhibitor?R115777in?adults?with?refractory?andrelapsed?acute?leukemias：a?phase?1clinical-laboratory?correlativetrial″Blood?97：3361-3369

Kohlmann?et?al.(2003)″Molecular?characterization?of?acuteleukemias?by?use?of?microarray?technology″Genes?Chrom.CancerGenes?Chrom.Cancer?37：396-405

Kurzrock?et?al.(2004)″Phase?II?study?of?R115777，a?farnesyltransferase?inhibitor，in?myelodysplastic?syndrome″J.Clin.Oncol.22：1287-1292

Lancet?et?al.(2003)″Farnesyltransferase?inhibitors?inhematologic?malignancies：new?horizons?in?therapy″Blood?102：3880-3889

Lancet?et?al.(2004)″Tipifarnib?(ZARNESTRA ^TM?in?PreviouslyUntreated?Poor-Risk?AML?of?the?Elderly：Updated?Results?of?aMulticenter?Phase?2Trial″Blood?104：874a

Li?et?al.(2002)″The?hematopoiesis-specific?GTP-binding?proteinRhoH?is?GTPase?deficient?and?modulates?activities?of?other?RhoGTPases?by?an?inhibitory?function″Mol.Cell.Biol.22：1158-1171

Lynch?et?al.(2004)″Activating?mutations?in?the?epidermalgrowth?factor?receptor?underlying?responsiveness?of?non-small-celllung?cancer?to?Gefitinib″N.Engl.J.Med.350：2129-2139

McLean?et?al.(2004)″Pharmacogenomic?analysis?of?cytogeneticresponse?in?chronic?myeloid?leukemia?patients?treated?with?imatinib″Clin.Cancer?Res.10：155-165

Methods?in?Molecular?Biology，Vol.33：In?Situ?HybridizationProtocols，Choo，ed.，Humana?Press，Totowa，NJ(1994)

Morgan?et?al.(2001)″Cell-cycle-dependent?activation?ofmitogen-activated?protein?kinase?kinase?(MEK-1/2)in?myeloidleukemia?cell?lines?and?induction?of?growth?inhibition?and?apoptosisby?inhibitors?of?RAS?signaling″Blood?97：1823-1834

Na?et?al.(2004)″Inhibition?of?farnesyltransferase?preventscollagen-induced?arthritis?by?down-regulation?of?inflammatory?geneexpression?through?suppression?of?p21(ras)-dependent?NF-kappaBactivation″J?Immunol.173：1276-1283.

Okutsu?et?al.(2002)″Prediction?of?chemosensitivity?for?patientswith?acute?myeloid?leukemia，according?to?expression?levels?of?28genes?selected?by?genome-wide?complementary?DNA?microarrayanalysis″Mol.Cancer?Ther.1：1035-1042

Paez?et?al.(2004)″EGFR?mutations?in?lung?cancer：correlationwith?clinical?response?to?gefitinib?therapy?activating?mutations?in?theepidermal?growth?factor?receptor?underlying?responsiveness?of?non-small-cell?lung?cancer?to?Gefitinib″Science?304：1497-1500

Pasqualucci?et?al.(2001)″Hypermutation?of?multiple?proto-oncogenes?in?B-cell?diffuse?large-cell?lymphomas″Nature?412：341-346

Pinkel(1988)″Fluorescence?in?situ?Hybridization?with?HumanChromosome-Specific?Libraries：Detection?of?Trisomy?21andTranslocations?of?Chromosome?4″Proc?Natl?Acad?Sci?USA?85：9138-9142

Pinkel?et?al.(1998)″High?resolution?analysis?of?DNA?copynumber?variation?using?comparative?genomic?hybridization?tomicroarrays″Nature?Genetics?20：207-211

Rao?et?al.(2004)″Phase?III?Double-Blind?Placebo-ControlledStudy?of?Farnesyl?Transferase?Inhibitor?R115777in?Patients?WithRefractory?Advanced?Colorectal?Cancer″J?Clin?Oncol?22：3950-3957

Raponi，et?al.(2004)″Identification?of?Molecular?Predictors?ofResponse?to?ZARNESTRA ^TM(Tipifarnib，R115777)in?Relapsed?andRefractory?Acute?Myeloid?Leukemia″Blood?104：861a

Reiss?et?al.(1990)″Inhibition?of?purified?p21ras?farnesyl：proteintransferase?by?Cys′AAX?tetrapeptides″Cell?62：81-88

Reuter?et?al.(2000)″Targeting?the?Ras?signaling?pathway：arational，mechanism-based?treatment?for?hematologic?malignancies？″Blood?96：1655-1669

Sahai?et?al.(2002)″RHO-GTPases?and?cancer″Nat.Rev.Cancer?2：133-142

Schoch?et?al.(2002)″Acute?myeloid?leukemias?with?reciprocalrearrangements?can?be?distinguished?by?specific?gene?expressionprofiles″Proc.Natl.Acad.Sci.USA?99：10008-10013

Sterpetti?et?al.(1999)″Activation?of?the?Lbc?Rho?exchangefactor?proto-oncogene?by?truncation?of?an?extended?C?terminus?thatregulates?transformation?and?targeting″Mol.Cell.Biol.19：1334-1345

Strachan?and?Read，Human?Molecular?Genetics，1996

Takada?et?al.(2004)″Protein?farnesyltransferase?inhibitor(SCH66336)abolishes?NF-kappaB?activation?induced?by?variouscarcinogens?and?inflammatory?stimuli?leading?to?suppression?of?NF-kappaB-regulated?gene?expression?and?up-regulation?of?apoptosis″JBiol?Chem.279：26287-26299.

Testa?et?al.(2004)″Interleukin-3receptor?in?acute?leukemia″Leukemia?18：219-226

Thomas?et?al.(2001)″R115777，a?farnesyl?transferase?inhibitor(FTI)，has?significant?anti-leukemia?activity?in?patients?with?chronicmyeloid?leukemia?(CML)″Blood?98

Tijssen(1993)Laboratory?Techniques?in?Biochemistry?andMolecular?Biology，Vol.24：Hybridization?With?Nucleic?Acid?Probes，Elsevier，NY

Toksoz?et?al.(1994)″Novel?human?oncogene?lbc?detected?bytransfection?with?distinct?homology?regions?to?signal?transductionproducts″Oncogene?9：621-628

Valk?et?al.(2004)″Prognostically?useful?gene-expression?profilesin?acute?myeloid?leukemia″N.Engl.J.Med.350：1617-1628

Van?Cutsem?et?al.(2004)″Phase?III?trial?of?gemcitabine?plustipifarnib?compared?with?gemcitabine?plus?placebo?in?advancedpancreatic?cancer″J.Clin.Oncol.22：1430-1438

Yagi?et?al.(2003)″Identification?of?a?gene?expression?signatureassociated?with?pediatric?AML?prognosis″Blood?102：1849-1856

Zhang?et?al.(2002)″Farnesyltransferase?inhibitors?reverse?Ras-mediated?inhibition?of?Fas?gene?expression″Cancer?Res.62：450-458

Zheng?et?al.(1995)″Direct?involvement?of?the?small?GTP-binding?protein?Rho?in?lbc?oncogene?function″J.Biol.Chem.270：9031-9034

Claims (39)

1. the method for assessment acute myelogenous leukemia (AML) state may further comprise the steps:

A. obtain biological sample from AML patient; And

B. measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, wherein above or the expression level that is lower than the described marker gene of predetermined threshold level indicate the AML state.

2. one kind to acute myelogenous leukemia (AML) patient method by stages, may further comprise the steps:

A. obtain biological sample from AML patient; And

B. measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, wherein above or the expression level that is lower than the described marker gene of predetermined threshold level indicate the AML survival rate.

3. the method for a definite acute myelogenous leukemia (AML) patient treatment scheme may further comprise the steps:

A. obtain biological sample from AML patient; And

B. measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, the expression level that wherein surpasses or be lower than the described marker gene of predetermined threshold level is enough to indicate replying treatment, make the doctor can determine the degree and the type of the treatment recommended, so that suitable treatment to be provided.

4. treatment acute myelogenous leukemia (AML) patient method may further comprise the steps:

A. obtain biological sample from AML patient; And

B. measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9, wherein surpass or the expression level that is lower than the described marker gene of predetermined threshold level indicates replying for the treatment of; And

If c. they have respondent's spectrum, treat described patient with assisting therapy.

5. the method for the high or low mortality ratio danger of a definite acute myelogenous leukemia (AML) patient may further comprise the steps:

A. obtain biological sample from AML patient; And

B. measure with corresponding to those the relevant biomarker of marker gene that is selected from table 3

The expression level that wherein surpasses or be lower than the described marker gene of predetermined threshold level is enough to indicate dead danger, so that the doctor can determine the degree and the type of the treatment recommended.

6. as each method of claim 1-5, also comprise the measurement at least a expression of gene level that express on composing type ground in described sample.

7. as each method of claim 1-5, wherein said specificity is at least about 40%.

8. as each method of claim 1-5, wherein said susceptibility is at least about 80%.

9. as the method for claim 28, wherein the P value is less than 0.05.

10. as each method of claim 1-5, wherein on microarray or gene chip, measure genetic expression.

11. as the method for claim 10, wherein said microarray is cDNA array or oligonucleotide array.

12. as the method for claim 10, wherein said microarray or gene chip also comprise mark reagent in one or more.

13. as each method of claim 1-5, wherein determine genetic expression by nucleic acid amplification, described nucleic acid amplification is by carrying out polymerase chain reaction (PCR) from the RNA of sample and realize extracting.

14. as the method for claim 13, wherein said PCR is reverse transcriptase polymerase chain reaction (RT-PCR).

15. as the method for claim 14, wherein said RT-PCR also comprises mark reagent in one or more.

16., wherein detect genetic expression by the albumen of measuring or detect described genes encoding as each method of claim 1-5.

17., wherein detect described albumen with being specific to described proteic antibody as the method for claim 16.

18., wherein detect genetic expression by the feature of measuring described gene as each method of claim 1-5.

19. as the method for claim 18, the feature of wherein said measurement is selected from DNA cloning, methylates, sudden change and allelic variation.

20. one kind generates acute myelogenous leukemia (AML) and prejudges patient's method of reporting, may further comprise the steps:

Determine the result of claim 1-5 in each; And

Prepare to show described result's report.

21. as the method for claim 20, wherein said report contains to patient's result and/or with respect to patient group's dangerous probability and/or likelihood or to the assessment of replying of chemotherapy.

22. patient's report that generates according to the method for claim 21.

23. in biological sample, measure to determine the test kit of acute myelogenous leukemia (AML) prognosis for one kind, comprise: be used to detect the isolated nucleic acid sequences of one group of gene, their complementary sequence or the material of its part, described gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

24., also comprise the reagent that is used to carry out microarray analysis as the test kit of claim 24.

25. as the test kit of claim 24, also comprise medium, described nucleotide sequence, they complementary sequence or its part by described medium and determined.

26. the goods of assessment acute myelogenous leukemia (AML) state, it comprises following substances, described material is used to detect the isolated nucleic acid sequences of one group of gene, their complementary sequence or its part, and described gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

27., also comprise the reagent that is used to carry out microarray analysis as the goods of claim 26.

28. as the goods of claim 27, also comprise medium, described nucleotide sequence, they complementary sequence or its part by described medium and determined.

29. one kind is used to carry out each the microarray or the gene chip of method of claim 1-5.

30. as the microarray of claim 29, comprise the isolated nucleic acid sequences of one group of gene, their complementary sequence or its part, described gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

31. as the microarray of claim 30, wherein said measurement or sign are that expression is expressed or owed at least 1.5 times of mistakes.

32. as the microarray of claim 30, wherein said measurement provided the statistically evident p value of expressing or owing to express.

33. as the microarray of claim 32, wherein the P value is less than 0.05.

34., comprise cDNA array or oligonucleotide array as the microarray of claim 30.

35., also comprise mark reagent in one or more as the microarray of claim 30.

36. a diagnosis/prognosis bag, it comprises the isolated nucleic acid sequences of one group of gene, their complementary sequence or its part, and described gene is selected from corresponding to those the marker gene that is selected from table 3, table 4, table 5, table 7, table 8 or the table 9.

37. as the bag of claim 36, wherein said measurement or be characterized by at least 1.5 times of mistakes and express or owe and express.

38. as the bag of claim 37, wherein said measurement provided the statistically evident p value of expressing or owing to express.

39. as the bag of claim 37, wherein the P value is less than 0.05.

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