CN103602746A - Biomarker for predicting neoadjuvant chemotherapy curative effect of breast cancer and application of biomarker - Google Patents
Biomarker for predicting neoadjuvant chemotherapy curative effect of breast cancer and application of biomarker Download PDFInfo
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Abstract
The invention discloses a biomarker for predicting neoadjuvant chemotherapy curative effect of breast cancer and application of the biomarker. The biomarker for predicting neoadjuvant chemotherapy curative effect of breast cancer is a long non-coding RNA and has a sequence shown in SEQ ID NO:1 or has a corresponding DNA sequence. The biomarker can be used for preparing a kit having one or more than one of functions of prediction of neoadjuvant chemotherapy curative effect of breast cancer, clinical diagnosis, drug screening and treatment assessment. The kit is an RT-qPCR kit or a sequencing kit for quantitatively or qualitatively detecting the long non-coding RNA. The application of the biomarker for predicting the neoadjuvant chemotherapy curative effect of breast cancer is clear, the difference (P is less than 0.05) of the biomarker in pCR group and NpCR group fresh breast cancer tissues is displayed, the expression in the NpCR group fresh breast cancer tissues is obviously higher than that in the pCR group, the biomarker can be used for predicting the neoadjuvant chemotherapy curative effect, and the blindness of the neoadjuvant chemotherapy is avoided.
Description
Technical field
The invention belongs to biotechnology and medical field, relate to a kind of biomarker of predicting neoadjuvant chemotherapy in breast curative effect and uses thereof.
Background technology
Mammary cancer is one of modal malignant tumour of women, and sickness rate accounts for the 7-10% of the various malignant tumours of whole body, occupies first of women's Cancer Mortality.Neoadjuvant chemotherapy in breast claims again preoperative chemotherapy or initial chemotherapy, essence belongs to adjuvant chemotherapy, but its possible mechanism of action is different from again general adjuvant chemotherapy of patients, it refers to the general that non-metastatic tumour is carried out before topical therapeutic, the treatment of systematic cytotoxic drug.At present, many Neoadjuvant Chemotherapies not only can be for advanced breast cancer patient, and can also allow increasing breast carcinoma of early stage patient be benefited, and has obtained good society and economic benefit.
New adjuvant chemotherapy has following 3 advantages to operable patient with breast cancer: (1) reaches Pathologic Complete and alleviates (pathologic complete remission, pCR), and patient's long-term survival rate is obviously improved.PCR is a level of signification as neoadjuvant chemotherapy in breast curative effect evaluation, according to the international Committee of Experts, recommend pCR to be defined as: after new adjuvant chemotherapy, in Postoperative Specimen at primary tumo(u)r tissue and axillary gland all without pernicious Histological Evidence, or only deposit carcinoma in situ composition.And after new adjuvant chemotherapy, as long as the former piece of swelling is or/and metastatic lesion still has infiltrating cancer tissue residue to be divided into non-pCR(Non-pathologic complete remission, NpCR) group.The median follow-up time was result of study of 16 years that NSABP B-18 tests up-to-date announcement shows: disease free survival rate (the disease-free survival of new adjuvant chemotherapy and adjuvant chemotherapy of patients, DFS) and overall survival (overall survival, OS) and no significant difference, but in new adjuvant chemotherapy group pCR patient's DFS and OS obviously more non-pCR patient is long.In addition, NSABP B-27 has compared pCR patient and the lifetime that has residual tumor patient after new adjuvant chemotherapy, finds that obviously extend the former lifetime, and HR is 0.33(P < 0.0001).(2) reduce neoplasm staging, local advanced breast cancer can improve resection rate after new adjuvant chemotherapy, and can increase the chance of protecting breast conserving surgery.NSABP B-18 result of study finds after new adjuvant chemotherapy that 67.8% patient obtains and protects newborn chance, and first operator protects newborn rate and is 59.8%, two group and has notable difference (P=0.002).(3) new adjuvant chemotherapy can carry out drug sensitive test in body, prevents resistance clone's appearance, for the selection of chemotherapy regimen provides reference frame.
Although, the clinical application history of existing more than 30 year of new adjuvant chemotherapy, obtained in recent years compared with much progress, but along with going deep into of research, also many problem demanding prompt solutions and shortcoming have been found: progression of disease (progression disease appears in the patient of (1) 8%-10% after accepting new adjuvant chemotherapy, PD), even lose the opportunity of topical therapeutic.(2) in addition, approximately 20% patient with breast cancer is insensitive to new adjuvant chemotherapy.For this part patient, if new adjuvant chemotherapy is insensitive the course for the treatment of long, also may incur loss through delay opportunity of topical therapeutic.These reasons, may make part doctor and patient too conservative when carrying out new adjuvant chemotherapy just, cause pCR to lead generally lower than 10%.PCR in B-18 and these two large-scale clinical trials of B-27 leads also only 13% and 26%.Therefore, need a kind of effective outcome prediction index, before chemotherapy, just can predict the curative effect of new adjuvant chemotherapy, avoid the blindness for the treatment of.
MALAT1 gene (metastasis associated in lung denocarcinoma transcript 1) is that human lung adenocarcinoma shifts associated retroviral book 1 gene, a new long non-coding RNA (the long non-coding RNA going out by subtractive hybridization Screening and Identification in the transfer of research Non-small cell lung carcinoma at first, LncRNA), it is to be positioned on mankind 11q13 karyomit(e), be about the non-coding RNA of 8 700 bp, lack significant open encoder block, cannot translate corresponding protein in vitro. be generally expressed in the histocyte of the mankind and mouse, especially outstanding with neural system.Nonsmall-cell lung cancer has in the cell strain of later stage metastasis tendency in early days, and its expression level is for significantly raising without metastasis tendency cell strain, with survival negative correlation.Many studies show that afterwards, the equal significant correlations of tissue malignancies such as MALAT1 and mammary gland, pancreas, lung, colon, prostate gland and liver.Research discovery, MALAT1 can regulate and control caspase (Caspase)-3 ,-8 in cervical cancer cell strain CaSKi apoptosis of tumor cells path, Ba, Bcl-2 and Bcl-xL etc., modulate tumor cell proliferation and invasion and attack.
Summary of the invention
The object of this invention is to provide a kind of biomarker of predicting neoadjuvant chemotherapy in breast curative effect and uses thereof.
Predict a biomarker for neoadjuvant chemotherapy in breast curative effect, it is a kind of long non-coding RNA, and sequence is as shown in SEQ ID NO:1, or its corresponding DNA sequence dna.
A purposes for described biomarker, the test kit of one or more in assessing for the preparation of prediction neoadjuvant chemotherapy in breast curative effect, clinical diagnosis, screening of medicaments, treatment.
Described test kit is RT-qPCR test kit or the sequencing kit of long non-coding RNA quantitatively or described in qualitative detection.
A kind of difference analysis method of described biomarker, flesh tissue before neoadjuvant chemotherapy in breast, pCR group and NpCR group are 15 examples, the RNA of pCR and NpCR extraction is mixed, carry out sequencing library structure, by the DNA library employing preparing while synthesizing sequencing carry out degree of depth order-checking, then order-checking Reads and human genome compare, by RPKM value, calculate, obtain the expression amount of each gene.
Beneficial effect of the present invention: the purposes of the biomarker of clear and definite prediction neoadjuvant chemotherapy in breast curative effect, neoadjuvant chemotherapy in breast pCR group and NpCR group gene expression difference are analyzed, result shows that this biomarker there are differences (P<0.05) in pCR group and NpCR organize fresh breast cancer tissue, and the expression of organizing in breast cancer tissue at NpCR is significantly higher than pCR group.Point out this biomarker can be used for predicting Efficacy of Neoadjuvant Chemotherapy, avoid new adjuvant chemotherapy blindness.
Accompanying drawing explanation
Fig. 1 is NpCR VS pCR MA Plot figure.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described further.
The gene M ALAT1 that the present invention finds is as follows for predicting the concrete grammar of neoadjuvant chemotherapy in breast curative effect:
One, experiment sample
Tumor tissues sample used in the present invention is from Zhejiang Prov. Tumor Hospital, and breast cancer tissue's sample is accepted the patient of new adjuvant chemotherapy from 30 examples, and pCR group and NpCR group are 15 routine patients.Coarse needle bioptic is transferred in centrifuge tube after getting tissue samples, puts into immediately liquid nitrogen cryopreservation, after to be stored in-80 ℃ of refrigerators stand-by.
Two, experiment reagent
Agents useful for same in RNA extraction and PCR product testing process, as acidic phenol (pH4.5), chloroform, Virahol, ethanol,
The routine biochemistry reagent such as DNA polyacrylamide gel electrophoresis reagent, are all purchased from Shanghai Sheng Gong company.Pcr amplification reagent (containing 10 * buffer, dNTPs mixture) and reverse transcription reaction agents useful for same, as dNTP mixture (each 10 mM for RNase inhibitor, M-MuLV ThermoScript II, 5 * M-MuLV damping fluid and reverse transcription, RNase-free), all purchased from TaKaRa company.Fluorescent quantificationally PCR detecting kit based on DNA dye method (containing real-time PCR Master Mix, 50 * ROX reference) is TOYOBO product.All primers entrust (miRNA loop-stem structure reverse transcription primer, PCR forward primer, PCR reverse primer) Shanghai biotechnology company limited synthetic.In RNA extraction and PCR product testing process, agents useful for same and buying are as listed in following table 1,2,3:
table 1 total RNA extracts agents useful for same and source thereof
table 2 miRNA reverse transcription agents useful for same and source thereof
table 3 real-time PCR agents useful for same and source thereof
Three, experimentation
first part: neoadjuvant chemotherapy in breast pCR group and NpCR group gene expression spectrum analysis
1, sample is selected
Enter group crowd for without distant metastasis, without whole body major disease, can surgical radical treatment infiltrative breast carcinoma patient totally 30 examples.Inclusive criteria is: can complete chemotherapy by predetermined scheme, II, III A phase tumour can obtain the total RNA of high quality of at least 10 ug from treating in frozen biopsy tissue.Clinical trial meets moral ethics regulation.The written signature clinical study of patient Informed Consent Form.
2, RNA extracts and high-throughput sequencing library structure
The RNA of pCR and NpCR extraction is mixed, carry out sequencing library structure, operation steps is as follows.
2.1 mRNA are separated
Get total RNA 10 ug that purifying is good, with Sera-mag Magnetic Oligo (dT) Beads(Thermo, part # 1004815) mRNA of decoupled band poly (A) tail from total RNA.
2.2 mRNA fragmentations
Fragmentation reaction system:
Place 2.5min for 94 ℃, place on ice immediately ethanol deposition and purification.
Synthesizing of 2.3 cDNA article one chains
Reaction system:
Place 5min, place 2min on ice for 65 ℃;
Add following reagent
Reaction conditions:
Place on ice.
Synthesizing of 2.4 cDNA second chains
In reaction product, add 51.5 ul NF-H
2o
Reaction system:
Place 5min on ice or until completely cooling;
Add following reagent:
Place 2.5h for 16 ℃;
With QIAquick PCR Purification Kit purified product, product is dissolved in 50 ul NF-H
2o.
2.5 cDNA ends are repaired
Reaction system:
Place 30min for 20 ℃;
With QIAquick PCR Purification Kit purified product, product is dissolved in 32 ul NF-H
2o.
2.6 3 ' the ends at DNA fragmentation add ' A '
Reaction system:
| Eluted DNA | 32 ul |
| 10 |
5 ul |
| 1 |
10 ul |
| Klenow Fragment (3’-5’ exo - minus) | 3 ul |
| Total | 50 ul |
Place 30min for 37 ℃;
With QIAquick PCR Purification Kit purified product, product is dissolved in 42 ul NF-H
2o.
2.7 cDNA ends connect
Reaction system:
| Eluted DNA | 42 ul |
| 10X T4 DNA ligase buffer with 10 |
5 ul |
| Adaptor mix (10 uM) | 1 ul |
| T4 DNA ligase (400 U/ul) | 2 ul |
| Total | 50 ul |
Place 2h for 23 ℃;
With QIAquick PCR Purification Kit purified product, product is dissolved in 10 ul NF-H
2o.
2.8 purifying cDNA templates
The sepharose of preparation 2%;
Click and enter sample, 50bp Marker, 75V, the about 1h of electrophoresis;
EB 20 min that dye;
Extract the smear region of 300bp-400bp;
With QIAquick Gel Extraction Kit purified product, product is dissolved in 50 ul NF-H
2o.
2.9 cDNA template amplifications
Reaction system
| NF-H2O | 3.5 ul |
| 5 |
10 ul |
| 2.5 |
5 ul |
| Multiplexing PCR Primer 1.0 | 2 ul |
| Multiplexing PCR Primer 2.0 | 2 ul |
| PCR Primer Index | 2 ul |
| cDNA | 25 ul |
| Phusion High-Fidelity DNA Polymerase (2 U/ul) | 0.5 ul |
| Total | 50 ul |
Reaction conditions
| 98 °C | 30 s |
| 98 °C |   |
| 65 °C | 30 s |
| 72 °C | 15 s |
| 72 °C | 7min |
| 8 °C | ∞ |
With Axygen PCR Clean Up Kit purified pcr product, product is dissolved in 50 ul NF-H
2o, gets 1ul and surveys OD value.
2.10 rubber tapping is reclaimed
The smear region cutting that is 300-350bp by size in sample is reclaimed, with QIAquick Gel Extraction Kit purified product.
2.11 Illumina Solexa degree of depth order-checkings
The DNA library preparing is diluted to 10nM with damping fluid, adopts while synthesizing sequencing (sequencing by synthesis, SBS), utilize Hiseq2000 to carry out two-way 1X50 nt order-checking, it is 50 bp that length is read in the order-checking of Read.
Sequencing result is analyzed
3.1 primitive sequencer Reads quality basic statistics are as shown in table 4 below:
Table 4
| Sample | NpCR group | PCR group |
| Total Reads | 13118494 | 10284940 |
| Length Reads | 50 | 50 |
| Q20 | 13099904 | 10269116 |
| Q20% | 99.86% | 99.85% |
| Q10 | 13118494 | 10284940 |
| Q10% | 100.00% | 100.00% |
Note: Q>20 (%)=Q>20 read/ Total read*100; The same Q20 of Q10.According to the technical documentation of Illumina, when mass value is 10, error rate is 10%, 20 to be 1%.
3.2 Reads quality pre-treatment
In view of the impact of sequencing data error rate on result, raw data is carried out to quality pre-treatment.
Quality pre-treatment step:
1) remove inferior quality Reads: quality threshold 20(error rate=1%), proportion threshold value 50%
2) remove in Reads containing N part
Before and after quality pre-treatment, data results statistical table is as follows:
3.3 gene expression spectrum analysis
Use SOAP software, Reads and the human genome of order-checking are compared, by RPKM value, calculate, obtain the expression amount of each gene.
RPKM be by map to the read number of gene divided by map the length (YiKBWei unit) to all read numbers (take million as unit) Yu the RNA of genome.
RNA-seq sees through the method that time technology of generation sequencing is detected gene performance amount, when weighing gene performance amount, if merely with map to read number calculate the performance amount of gene, in statistics, be one and quite do not conform to director, because the in the situation that of stochastic sampling, the probability that the gene that sequence is longer is pumped to originally will be shorter than sequence gene higher, thus, it is higher that the gene always that sequence is long is considered to performance amount, and misestimate the real performance amount of gene, so the people such as Ali Mortazavi proposed with RPKM in the performance amount of estimating gene in 2008.
Its formula is:
The gene expression abundance of this gene of DEGseq software analysis in different samples.Differential expression analysis relatively, wherein we choose | log2 (fold_change) | the gene of >1 and P_value<0.05 is as the screening conditions of differential expression.
As shown in Figure 1, NpCR and pCR group have the gene that significant difference is expressed to represent (being positioned at the upper right corner and the lower right corner that asymptotic dotted line surrounds) with point; We analyze annotated 33121 genes, with | log2 (fold_change) | the gene of >1 and P_value<0.05, as the screening conditions of differential expression, obtains the gene of 710 differential expressions altogether.Application RT-qPCR technology is carried out check analysis to MALAT1, further confirms that MALAT1 there are differences in pCR group and NpCR organize fresh breast cancer tissue.
SEQUENCE LISTING
<110> Zhejiang Prov. Tumor Hospital
Biomarker of <120> prediction neoadjuvant chemotherapy in breast curative effect and uses thereof
<160> 1
<170> PatentIn version 3.5
<210> 1
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<213> Homo sapiens
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Claims (4)
1. a biomarker of predicting neoadjuvant chemotherapy in breast curative effect, is characterized in that, it is a kind of long non-coding RNA, and sequence is as shown in SEQ ID NO:1, or its corresponding DNA sequence dna.
2. a purposes for the biomarker as described in claim 1, is characterized in that, the test kit of one or more in assessing for the preparation of prediction neoadjuvant chemotherapy in breast curative effect, clinical diagnosis, screening of medicaments, treatment.
3. the purposes of the biomarker as described in claim 1, is characterized in that, described test kit is RT-qPCR test kit or the sequencing kit of long non-coding RNA quantitatively or described in qualitative detection.
4. the difference analysis method of a biomarker as claimed in claim 1, it is characterized in that, flesh tissue before neoadjuvant chemotherapy in breast, pCR group and NpCR group are 15 examples, and the RNA of pCR and NpCR extraction is mixed, and carry out sequencing library structure, by the DNA library employing preparing while synthesizing sequencing carry out degree of depth order-checking, then Reads and the human genome of order-checking are compared, by RPKM value, calculate, obtain the expression amount of each gene.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105039319A (en) * | 2015-06-22 | 2015-11-11 | 浙江理工大学 | Biomarker for forecasting new auxiliary chemotherapy curative effect of breast cancer and fluorescence quantitative immune PCR kit |
| CN106550605A (en) * | 2014-03-18 | 2017-03-29 | 汉诺威医学院 | For the mitochondria non-coding RNA of progression of disease in predicting heart failure and Patients With Myocardial Infarction |
| CN107502671A (en) * | 2017-10-12 | 2017-12-22 | 中国医学科学院肿瘤医院 | Herceptin lower rectal cancer patient with breast cancer curative effect reagent box and its application are predicted using TRBV family members |
| CN108315430A (en) * | 2018-04-28 | 2018-07-24 | 上海交通大学医学院附属仁济医院 | Purposes of the PDL1 SNP genotype as prediction neoadjuvant chemotherapy in breast curative effect marker |
| CN110684847A (en) * | 2019-04-15 | 2020-01-14 | 德阳市人民医院 | Application of biomarker related to breast cancer occurrence and development |
| CN110923321A (en) * | 2019-12-26 | 2020-03-27 | 广东省人民医院(广东省医学科学院) | Annular RNA detection kit for predicting neoadjuvant chemotherapy reactivity of triple-negative breast cancer |
| CN117038010A (en) * | 2023-10-10 | 2023-11-10 | 中国人民解放军总医院 | Treatment image generation method and system in chemotherapy preparation stage |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101258250A (en) * | 2005-07-07 | 2008-09-03 | 拜奥默里克斯股份有限公司 | Method for breast cancer diagnosis |
| WO2013134558A1 (en) * | 2012-03-07 | 2013-09-12 | The Texas A & M University System | Cancer treatment targeting non-coding rna overexpression |
-
2013
- 2013-11-26 CN CN201310605932.4A patent/CN103602746A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101258250A (en) * | 2005-07-07 | 2008-09-03 | 拜奥默里克斯股份有限公司 | Method for breast cancer diagnosis |
| WO2013134558A1 (en) * | 2012-03-07 | 2013-09-12 | The Texas A & M University System | Cancer treatment targeting non-coding rna overexpression |
Non-Patent Citations (4)
| Title |
|---|
| LARS HENNING SCHMIDT等: "The Long Noncoding MALAT-1 RNA Indicates a Poor Prognosis in Non-small Cell Lung Cancer and Induces Migration and Tumor Growth", 《JOURNAL OF THORACIC ONCOLOGY》 * |
| YANG,F等: "NR_002819.2", 《GENBANK》 * |
| 宋皓军等: "长链非编码RNA与肿瘤的关系及其临床价值", 《中国细胞生物学学报》 * |
| 江艳等: "MALAT-1与肿瘤", 《生命的化学》 * |
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|---|---|---|---|---|
| CN106550605A (en) * | 2014-03-18 | 2017-03-29 | 汉诺威医学院 | For the mitochondria non-coding RNA of progression of disease in predicting heart failure and Patients With Myocardial Infarction |
| CN105039319A (en) * | 2015-06-22 | 2015-11-11 | 浙江理工大学 | Biomarker for forecasting new auxiliary chemotherapy curative effect of breast cancer and fluorescence quantitative immune PCR kit |
| CN107502671A (en) * | 2017-10-12 | 2017-12-22 | 中国医学科学院肿瘤医院 | Herceptin lower rectal cancer patient with breast cancer curative effect reagent box and its application are predicted using TRBV family members |
| CN107502671B (en) * | 2017-10-12 | 2020-05-05 | 中国医学科学院肿瘤医院 | Kit for predicting curative effect of trastuzumab neoadjuvant therapy of breast cancer patient by using TRBV family member and application thereof |
| CN108315430A (en) * | 2018-04-28 | 2018-07-24 | 上海交通大学医学院附属仁济医院 | Purposes of the PDL1 SNP genotype as prediction neoadjuvant chemotherapy in breast curative effect marker |
| CN108315430B (en) * | 2018-04-28 | 2021-07-23 | 上海交通大学医学院附属仁济医院 | Application of PDL1SNP genotype as marker for predicting neoadjuvant chemotherapy curative effect of breast cancer |
| CN110684847A (en) * | 2019-04-15 | 2020-01-14 | 德阳市人民医院 | Application of biomarker related to breast cancer occurrence and development |
| CN110684847B (en) * | 2019-04-15 | 2020-11-06 | 德阳市人民医院 | Application of biomarker related to breast cancer occurrence and development |
| CN110923321A (en) * | 2019-12-26 | 2020-03-27 | 广东省人民医院(广东省医学科学院) | Annular RNA detection kit for predicting neoadjuvant chemotherapy reactivity of triple-negative breast cancer |
| CN117038010A (en) * | 2023-10-10 | 2023-11-10 | 中国人民解放军总医院 | Treatment image generation method and system in chemotherapy preparation stage |
| CN117038010B (en) * | 2023-10-10 | 2024-02-13 | 中国人民解放军总医院 | Treatment image generation method and system in chemotherapy preparation stage |
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Application publication date: 20140226 |