CN107746887A - LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer transfering reagent boxes - Google Patents
LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer transfering reagent boxes Download PDFInfo
- Publication number
- CN107746887A CN107746887A CN201711142657.1A CN201711142657A CN107746887A CN 107746887 A CN107746887 A CN 107746887A CN 201711142657 A CN201711142657 A CN 201711142657A CN 107746887 A CN107746887 A CN 107746887A
- Authority
- CN
- China
- Prior art keywords
- lncrna
- breast cancer
- bone
- qpcr
- sequence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 206010006187 Breast cancer Diseases 0.000 title claims abstract description 178
- 208000026310 Breast neoplasm Diseases 0.000 title claims abstract description 177
- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 92
- 238000003745 diagnosis Methods 0.000 title claims abstract description 83
- 208000026535 luminal A breast carcinoma Diseases 0.000 title claims abstract description 37
- 239000000203 mixture Substances 0.000 title claims abstract description 17
- 108091046869 Telomeric non-coding RNA Proteins 0.000 title claims description 33
- 239000003153 chemical reaction reagent Substances 0.000 title abstract description 11
- 108020005198 Long Noncoding RNA Proteins 0.000 claims abstract description 222
- 238000012546 transfer Methods 0.000 claims abstract description 87
- 206010005949 Bone cancer Diseases 0.000 claims abstract description 52
- 210000002966 serum Anatomy 0.000 claims abstract description 50
- 238000011529 RT qPCR Methods 0.000 claims description 99
- 230000014509 gene expression Effects 0.000 claims description 72
- 238000009007 Diagnostic Kit Methods 0.000 claims description 41
- 230000000692 anti-sense effect Effects 0.000 claims description 31
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 claims description 27
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 claims description 27
- 238000000034 method Methods 0.000 claims description 21
- 238000011144 upstream manufacturing Methods 0.000 claims description 16
- 238000007477 logistic regression Methods 0.000 claims description 14
- 102000004190 Enzymes Human genes 0.000 claims description 9
- 108090000790 Enzymes Proteins 0.000 claims description 9
- 210000004369 blood Anatomy 0.000 claims description 7
- 230000015271 coagulation Effects 0.000 claims description 6
- 238000005345 coagulation Methods 0.000 claims description 6
- 210000003462 vein Anatomy 0.000 claims description 6
- 101100314454 Caenorhabditis elegans tra-1 gene Proteins 0.000 abstract description 34
- 230000002018 overexpression Effects 0.000 abstract description 33
- 238000012360 testing method Methods 0.000 abstract description 33
- 108091007417 HOX transcript antisense RNA Proteins 0.000 abstract description 26
- 102100039540 Exocyst complex component 7 Human genes 0.000 abstract description 25
- 101000813489 Homo sapiens Exocyst complex component 7 Proteins 0.000 abstract description 25
- 238000002360 preparation method Methods 0.000 abstract description 2
- 102100027341 Neutral and basic amino acid transport protein rBAT Human genes 0.000 abstract 1
- 101710159376 Neutral and basic amino acid transport protein rBAT Proteins 0.000 abstract 1
- 238000002405 diagnostic procedure Methods 0.000 description 52
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 28
- 108020004414 DNA Proteins 0.000 description 20
- 238000011156 evaluation Methods 0.000 description 16
- 239000012141 concentrate Substances 0.000 description 13
- 206010028980 Neoplasm Diseases 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 12
- 230000035945 sensitivity Effects 0.000 description 12
- 230000004069 differentiation Effects 0.000 description 11
- 238000013211 curve analysis Methods 0.000 description 9
- 201000011510 cancer Diseases 0.000 description 8
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 206010027476 Metastases Diseases 0.000 description 7
- 238000002591 computed tomography Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 239000012807 PCR reagent Substances 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 238000013480 data collection Methods 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000001962 electrophoresis Methods 0.000 description 4
- 230000009401 metastasis Effects 0.000 description 4
- 108091027963 non-coding RNA Proteins 0.000 description 4
- 102000042567 non-coding RNA Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000010839 reverse transcription Methods 0.000 description 4
- 238000012109 statistical procedure Methods 0.000 description 4
- 208000003721 Triple Negative Breast Neoplasms Diseases 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 208000022679 triple-negative breast carcinoma Diseases 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- -1 ER) Proteins 0.000 description 2
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 108010029485 Protein Isoforms Proteins 0.000 description 2
- 102000001708 Protein Isoforms Human genes 0.000 description 2
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 2
- 240000002825 Solanum vestissimum Species 0.000 description 2
- 235000018259 Solanum vestissimum Nutrition 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000007469 bone scintigraphy Methods 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 102000003998 progesterone receptors Human genes 0.000 description 2
- 108090000468 progesterone receptors Proteins 0.000 description 2
- 238000004393 prognosis Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 241000023308 Acca Species 0.000 description 1
- 241000208340 Araliaceae Species 0.000 description 1
- 206010055113 Breast cancer metastatic Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102100038595 Estrogen receptor Human genes 0.000 description 1
- 101000851181 Homo sapiens Epidermal growth factor receptor Proteins 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000006718 epigenetic regulation Effects 0.000 description 1
- 108010038795 estrogen receptors Proteins 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000027706 hormone receptor-positive breast cancer Diseases 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000025915 regulation of apoptotic process Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Hospice & Palliative Care (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses the purposes of lncRNA compositions and preparation diagnosis indication LuminalA type Bone of Breast Cancer transfering reagent boxes.Present invention discover that, serum lncRNAXLOC_004122, SUMO1P3 and NBAT 1 can combine for diagnosing the Bone of Breast Cancer transfer of indication Luminal A types, serum lncRNAXLOC_004122, Linc00467 and lncRNAAl049452 can combine for diagnosing the Bone of Breast Cancer transfer of indication Luminal Type Bs, serum lncRNAAK043773 and EXOC7 can combine for diagnosing indication Her 2 overexpression type Bone of Breast Cancer transfers, lncRNA Lnc01089 and HOTAIR can combine for diagnosing three negative type breast cancers Bone tumours of indication, degree of accuracy height (more than 90%), testing cost is low, non-invasi, it is convenient and swift.
Description
Technical field
The invention belongs to biochemical field, is related to diagnosis composition and diagnostic kit, and in particular to a kind of lncRNA
Diagnosis composition and the application in terms of the diagnostic kit for diagnosing the Bone of Breast Cancer transfer of indication different molecular hypotype is prepared.
Background technology
Breast cancer is one of malignant tumour for threatening women life and health, there are about the people of 40-45 ten thousand every year and dies from breast cancer (ginseng
Examine document:Different molecular hypotype Bone of Breast Cancer shifts the Clinical symptoms and prognostic analysis of patient, XI AN JIAOTONG UNIVERSITY Subject Index doctor
Learn version, in September, 2017 the 5th phase of volume 38).Breast cancer is very easy to that DISTANT METASTASES IN occurs, and bone is the most common distant place of breast cancer
Metastasis site, it is bone tissue (bibliography more than the starting metastasis site of 50% patient:Genes associated with
breast cancer metastatic to bone,J Clin Oncol,2006;Implications of Bone-Only
Metastases in Breast Cancer:Favorable Preference with Excellent Outcomes of
Hormone Receptor Positive Breast Cancer,CancerRes Treat,2011).According to ERs
(estrogen receptor, ER), progesterone receptor (progesterone receptor, PR), human epidermal growth factor receptor
Breast cancer, can be divided into by the expression of body -2 (human epidermal growth factor receptor-2, HER-2)
4 hypotypes, it is respectively:Luminal A types, Luminal Type Bs, Her-2 overexpressions type and triple negative breast cancer (bibliography:
Gene expression patterns ofbreast carcinomas distinguish tumor subclasses
with clinical implications,PNAS,2001).Research shows that prognosis and its molecule of Bone of Breast Cancer transfer divide
Closely related (the bibliography such as type, clinical stages, lymph node status:Prevalence and risk factors ofbone
metastasis and skeletal related events in patients with primary breast cancer
in Japan,Int J Clin Onco,2014).The specific molecular biology of different molecular hypotype breast cancer and clinical pathology are special
Sign, determine the difference of its therapeutic modality and prognosis.Different molecular hypotype Bone of Breast Cancer transfer gene expression dose there is also
Difference.
Early diagnosis Bone of Breast Cancer transfer is to save the key of patient vitals.At present, radionuclide bone scan (ECT),
CT scan (CT), nuclear magnetic resonance (MRI), Positron emission computed tomography (PET-CT) and bone tissue
Biopsy is to find and make a definite diagnosis the goldstandard of Bone of Breast Cancer transfer.But there is different deficiencies, such as Laboratory Fee in these methods
With height, intervention diagnosis adds the burden of patient.Which increase the pressure of patient with breast cancer's Bone tumour conventional detection.
Long-chain non-coding RNA (long non-coding RNA, lncRNA) refers to more than 200 nucleotides of length, had
The non-coding RNA of controlling gene expressional function.Recent study shows, generation, evolution of the long-chain non-coding RNA in tumour
In play rush cancer or cancer suppressing action, they take part in apoptosis regulation, it is tumor-infiltrated with transfer etc. process;In addition, they
The growth of tumour cell is also influenceed by way of epigenetic regulation, promises to be novel tumor markers and oncotherapy
Target spot, good potential applicability in clinical practice (bibliography is shown in terms of tumor diagnosis and therapy:Long-chain non-coding RNA with
The relation and its clinical value of tumour, Chinese cell biology journal, the 7th phase of volume 34 in 2012).
Research shows that the several genes expression of breast cancer primary tumo(u)r is necessary to Bone tumour occurs, it is thus regarded that special
The transfer for determining organ is the coefficient result of multiple-factor, conclusion prompting, and the transspecific of bone is bone in primary tumo(u)r
The selection of different phenotype tumour cells and the result of bone source sex factor induction, the difference table of different molecular hypotype Bone of Breast Cancer transfer
It is expected to turn into the diagnosis marker (bibliography of Bone of Breast Cancer transfer up to gene:KangY,Siegel PM,ShuW,et
al.Amultigenic program mediatingbreast cancer metastasis to bone.Cancer Cell,
2003)。
Applicant is intended to research and compares lncRNA tables in generation Bone tumour and the blood serum of patients with human breast carcinoma that Bone tumour does not occur
The difference reached, find, verify the lncRNA marks that may be used as diagnosing, indicate the transfer of different molecular hypotype Bone of Breast Cancer, with
There is provided it is a kind of can the kit that shifts of quick diagnosis, indication different molecular hypotype Bone of Breast Cancer and method by blood.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of lncRNA diagnosis compositions, one is prepared into
Kind testing cost is low, non-invasi, the diagnostic kit of convenient and swift diagnosis indication different molecular hypotype Bone of Breast Cancer transfer.
The above-mentioned purpose of the present invention is achieved by following technical scheme:
First, Luminal A types Bone of Breast Cancer shifts
A kind of lncRNA diagnosis compositions, by lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA
NBAT-1 is formed.
Above-mentioned diagnosis composition is in terms of the diagnostic kit for diagnosing the Bone of Breast Cancer transfer of indication Luminal A types is prepared
Using.
For diagnosing the diagnostic kit of indication Luminal A types Bone of Breast Cancer transfer, including lncRNA XLOC_
004122nd, lncRNA SUMO1P3 and lncRNANBAT-1 qPCR primers.
Preferably, in described diagnostic kit, lncRNA XLOC_004122 qPCR sense primers such as Sequence
Shown in NO.1, qPCR anti-sense primers are as shown in Sequence NO.2.
Preferably, in described diagnostic kit, lncRNA SUMO1P3 qPCR sense primers such as Sequence NO.3
Shown, qPCR anti-sense primers are as shown in Sequence NO.4.
Preferably, in described diagnostic kit, lncRNA NBAT-1 qPCR sense primers such as Sequence NO.5
Shown, qPCR anti-sense primers are as shown in Sequence NO.6.
Preferably, internal reference GAPDH qPCR primers are also included in described diagnostic kit.
Preferably, in described diagnostic kit, the qPCR sense primers such as Sequence NO.19 of the internal reference GAPDH
Shown, qPCR anti-sense primers are as shown in Sequence NO.20.
Also include the enzyme needed for qRT-PCR in any of the above-described described diagnostic kit.
A kind of method for diagnosing the Bone of Breast Cancer transfer of indication Luminal A types, comprises the following steps:
Step S1, Luminal A type patient with breast cancer's limosis vein bloods are gathered, serum is centrifuged out after natural coagulation;
Step S2, serum total serum IgE is extracted, lncRNA XLOC_004122, lncRNA in total serum IgE are determined with qRT-PCR methods
SUMO1P3 and lncRNA NBAT-1 use X successively relative to internal reference GAPDH relative expression levels3、X1、X2Represent;
Step S3, by X3、X1、X2Value substitutes into dualistic logistic regression equation Y=-2.577+2.045X1+1.956X2+
1.676X3Y value is obtained, Y value is more than 0.598 and indicates that Bone tumour occurs for the patient with breast cancer, and bone does not occur less than 0.598 indication turns
Move.
2nd, Luminal Type Bs Bone of Breast Cancer shifts
A kind of lncRNA diagnosis compositions, by lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA
Al049452 is formed.
Above-mentioned diagnosis composition is in terms of the diagnostic kit for diagnosing the Bone of Breast Cancer transfer of indication Luminal Type Bs is prepared
Using.
For diagnosing the diagnostic kit of indication Luminal Type Bs Bone of Breast Cancer transfer, including lncRNA XLOC_
004122nd, lncRNA Linc00467 and lncRNA Al049452 qPCR primers.
Preferably, in described diagnostic kit, lncRNA XLOC_004122 qPCR sense primers such as Sequence
Shown in NO.1, qPCR anti-sense primers are as shown in Sequence NO.2.
Preferably, in described diagnostic kit, lncRNA Linc00467 qPCR sense primers such as Sequence
Shown in NO.7, qPCR anti-sense primers are as shown in Sequence NO.8.
Preferably, in described diagnostic kit, lncRNA Al049452 qPCR sense primers such as Sequence
Shown in NO.9, qPCR anti-sense primers are as shown in Sequence NO.10.
Preferably, in described diagnostic kit, internal reference GAPDH qPCR primers are included.
Preferably, in described diagnostic kit, the qPCR sense primers such as Sequence NO.19 of the internal reference GAPDH
Shown, qPCR anti-sense primers are as shown in Sequence NO.20.
Any of the above-described described diagnostic kit also includes the enzyme needed for qRT-PCR.
A kind of method for diagnosing the Bone of Breast Cancer transfer of indication Luminal Type Bs, comprises the following steps:
Step S1, Luminal Type B patient with breast cancer's limosis vein bloods are gathered, serum is centrifuged out after natural coagulation;
Step S2, serum total serum IgE is extracted, lncRNA XLOC_004122, lncRNA in total serum IgE are determined with qRT-PCR methods
Linc00467 and lncRNA Al049452 use X relative to internal reference GAPDH relative expression levels3、X1、X2Represent;
Step S3, by X3、X1、X2Value substitutes into dualistic logistic regression equation Y=Y=-2.241+1.883X1+2.275X2+
1.975X3Y value is obtained, Y value is more than 0.607 and indicates that Bone tumour occurs for the patient with breast cancer, and bone does not occur less than 0.607 indication turns
Move.
3rd, Her-2 overexpressions type Bone of Breast Cancer shifts
A kind of lncRNA diagnosis compositions, are made up of lncRNAAK043773 and lncRNAEXOC7.
Above-mentioned diagnosis composition is in terms of the diagnostic kit for diagnosing the Bone of Breast Cancer transfer of indication Her-2 overexpression types is prepared
Application.
A kind of diagnostic kit for being used to diagnose the Bone of Breast Cancer transfer of indication Her-2 overexpression types, including lncRNA
AK043773 and lncRNA EXOC7 qPCR primers.
Preferably, in described diagnostic kit, lncRNA AK043773 qPCR sense primers such as Sequence
Shown in NO.11, qPCR anti-sense primers are as shown in Sequence NO.12.
Preferably, in described diagnostic kit, lncRNA EXOC7 qPCR sense primers such as Sequence NO.13
Shown, qPCR anti-sense primers are as shown in Sequence NO.14.
Preferably, in described diagnostic kit, internal reference GAPDH qPCR primers are included.
Preferably, in described diagnostic kit, the qPCR sense primers such as Sequence NO.19 of the internal reference GAPDH
Shown, qPCR anti-sense primers are as shown in Sequence NO.20.
Any of the above-described described diagnostic kit also includes the enzyme needed for qRT-PCR.
A kind of method for diagnosing the Bone of Breast Cancer transfer of indication Her-2 overexpression types, comprises the following steps:
Step S1, Her-2 overexpression type patient with breast cancer's limosis vein bloods are gathered, bleeding is centrifuged after natural coagulation
Clearly;
Step S2, serum total serum IgE is extracted, lncRNA AK043773 and lncRNA in total serum IgE are determined with qRT-PCR methods
EXOC7 uses X successively relative to internal reference GAPDH relative expression levels1、X2Represent;
Step S3, by X1、X2Value substitutes into dualistic logistic regression equation Y=-2.918+2.618X1+2.115X2Obtain Y value, Y
Value is more than 0.495 and indicates that Bone tumour occurs for the patient with breast cancer, and Bone tumour does not occur less than 0.495 indication.
4th, triple negative breast cancer Bone tumour
A kind of lncRNA diagnosis compositions, it is made up of lncRNA Lnc01089 and lncRNA HOTAIR.
Above-mentioned diagnosis composition answering in terms of the diagnostic kit for diagnosing three negative type breast cancers Bone tumours of indication is prepared
With.
A kind of diagnostic kit for being used to diagnose three negative type breast cancers Bone tumours of indication, including lncRNA Lnc01089
With lncRNA HOTAIR qPCR primers.
Preferably, in described diagnostic kit, lncRNA Lnc01089 qPCR sense primers such as Sequence
Shown in NO.15, qPCR anti-sense primers are as shown in Sequence NO.16.
Preferably, in described diagnostic kit, lncRNA HOTAIR qPCR sense primers such as Sequence NO.17
Shown, qPCR anti-sense primers are as shown in Sequence NO.18.
Preferably, in described diagnostic kit, internal reference GAPDH qPCR primers are included.
Preferably, in described diagnostic kit, the qPCR sense primers such as Sequence NO.19 of the internal reference GAPDH
Shown, qPCR anti-sense primers are as shown in Sequence NO.20.
Any of the above-described described diagnostic kit also includes the enzyme needed for qRT-PCR.
A kind of method for diagnosing three negative type breast cancers Bone tumours of indication, comprises the following steps:
Step S1, three negative type breast cancers patient's limosis vein bloods are gathered, serum is centrifuged out after natural coagulation;
Step S2, serum total serum IgE is extracted, lncRNA Lnc01089 and lncRNA in total serum IgE are determined with qRT-PCR methods
HOTAIR uses X successively relative to internal reference GAPDH relative expression levels1、X2Represent;
Step S3, by X1、X2Value substitutes into dualistic logistic regression equation Y=-2.537+2.793X1+2.181X2Obtain Y value, Y
Value is more than 0.633 and indicates that Bone tumour occurs for the patient with breast cancer, and Bone tumour does not occur less than 0.633 indication.
It is a discovery of the invention that serum lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 can join
Share in diagnosis indication Luminal A types breast cancer whether Bone tumour, concentrate diagnosis indication rate of accuracy reached 90% in individual authentication
More than;Serum lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 can combine for diagnosing
Indicate Luminal Type Bs breast cancer whether Bone tumour, concentrate diagnosis indication rate of accuracy reached more than 90% in individual authentication;Serum
LncRNA AK043773 and lncRNA EXOC7 can combine whether bone turns for diagnosing indication Her-2 overexpression types breast cancer
Move, diagnosis indication rate of accuracy reached more than 90% is concentrated in individual authentication;LncRNA Lnc01089 and lncRNA HOTAIR can be with
Combine for diagnose indication three negative type breast cancers whether Bone tumour, individual authentication concentrate diagnosis indication rate of accuracy reached 90% with
On.Diagnosing the transfer of indication different molecular hypotype Bone of Breast Cancer using above-mentioned serum lncRNA, not only the degree of accuracy is high, and detect into
This low, non-invasi, convenient and swift, very big reduction patient suffering and burden.
Brief description of the drawings
Fig. 1 is that lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 combine and be used in test set
Diagnosis distinguishes Luminal A types breast cancer and does not shift the ROC curve shifted with Luminal A types Bone of Breast Cancer;
Fig. 2 combines for checking concentration lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 and is used for
Diagnosis distinguishes Luminal A types breast cancer and does not shift the accuracy rate shifted with Luminal A types Bone of Breast Cancer;
Fig. 3 is lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 joints in test set
The ROC curve shifted with Luminal Type Bs Bone of Breast Cancer is not shifted for diagnosing differentiation Luminal Type Bs breast cancer;
Fig. 4 concentrates lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 joints for checking
The accuracy rate shifted with Luminal Type Bs Bone of Breast Cancer is not shifted for diagnosing differentiation Luminal Type Bs breast cancer;
Fig. 5 is that lncRNA AK043773 and lncRNA EXOC7 combine for diagnosing differentiation Her-2 overexpressions in test set
Type breast cancer does not shift the ROC curve with the transfer of Her-2 overexpression types Bone of Breast Cancer;
Fig. 6 distinguishes Her-2 overexpressions to verify that concentration lncRNA AK043773 and lncRNA EXOC7 combines for diagnosing
Type breast cancer does not shift the accuracy rate with the transfer of Her-2 overexpression types Bone of Breast Cancer;
Fig. 7 is that lncRNA Lnc01089 and lncRNA HOTAIR combine for diagnosing differentiation three negative types breast in test set
Gland cancer does not shift the ROC curve with three negative type breast cancers Bone tumours;
Fig. 8 concentrates lncRNA Lnc01089 and lncRNA HOTAIR Combining diagnosis to distinguish three negative type breast cancers for checking
The accuracy rate with three negative type breast cancers Bone tumours is not shifted.
Embodiment
Essentiality content of the present invention is specifically introduced with reference to the accompanying drawings and examples, but the guarantor of the present invention is not limited with this
Protect scope.
All breast cancer samples of this project are taken from September, 2014 to 2017 Nian9Yue Lai Hospital Attached to Nantong Univ. or south
Tong Shi First People's Hospital or Nanjing drum tower hospital inspection are diagnosed as the patient of other malignant tumours of breast cancer and nonjoinder.It is all
Sample is divided into Luminal A types, Luminal Type Bs, Her-2 overexpressions type and three negative types according to SABC detection, various
Molecular isoform is according to whether transfer is divided into the non-transfer group of breast cancer and Bone of Breast Cancer transfer group, and each case of Bone of Breast Cancer transfer group
It is starting DISTANT METASTASES IN position to belong to bone.The non-transfer group of breast cancer and Bone of Breast Cancer transfer group pass through radionuclide bone scan
(ECT), CT scan (CT), nuclear magnetic resonance (MRI), Positron emission computed tomography (PET-CT)
And/or the inspection such as tissue biopsy confirms.The non-transfer group of breast cancer and Bone tumour group patient age compare without bright in each molecular isoform
Significant difference is different, has comparativity.Each group sample is finally half-and-half divided into test set at random and checking collects.
All sample packet information and sample number are as shown in the table after the diagnosis of above-mentioned goldstandard:
The collection of serum specimen:Patient limosis vein blood 5.0mL is gathered, centrifuged after natural coagulation (4000r/min, 2860
× g) serum is isolated after 7min, -80 DEG C of preservations are placed in, for detecting the relative expression quantity of target lncRNA in serum.
Embodiment 1:Luminal A types Bone of Breast Cancer shifts
First, experiment sample and experimental method
1st, experiment sample
The non-transfer group of breast cancer, the test set of Luminal A type Bone of Breast Cancer transfer groups and checking in Luminal A types
Collection.
2nd, RNA extractings and qRT-PCR
Total serum IgE is extracted from serum sample using Trizol reagents (Invitrogen, Chinese Shanghai).Use NanoDrop
ND-2000 spectrophotometers (Thermo Scientific) determine the concentration and purity of total serum IgE at 260nm, and electrophoresis detection shows
Show, the RNA mass of purification well carries out subsequent operation afterwards.Total serum IgE is turned using reverse transcription reagent box (Takara, DaLian, China)
Turn to cDNA.Quantitative fluorescent PCR (Takara, DaLian, China), and application ABI Prism7000 are carried out using SYBR Green methods
Quantitative fluorescent PCR system (Agilent Technologies) carries out Data Collection.Primer is closed by winning still biological (Chinese Shanghai)
Into lncRNA XLOC_004122 primer sequences:- the CTGGCAGGAACACCGGGTACTT-3 ' of upstream 5 ', downstream 5 '-
TGACTTTTACTTAGGAGCCACTTCTTG-3’;LncRNA SUMO1P3 primer sequences:Upstream 5 '-
CTGGAACTGGGAATGGAGGAAGA-3 ' ,-GATTGAGAAAGGATTGAGGGAAA-3 ' of downstream 5 ';LncRNA NBAT-1 draw
Thing sequence:- the CTGGGAAAGCCTGTGCTCTTGGA-3 ' of upstream 5 ' ,-GCTTCACAGTGCTGCTCAATCGT-3 ' of downstream 5 ';
GAPDH primer sequences:- the CGCTCTCTGCTCCTCCTGTTC-3 ' of upstream 5 ' ,-ATCCGTTGACTCCGACCTTCAC- of downstream 5 '
3’.3 average values measured are taken with 2-ΔΔCtMethod calculates lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA
NBAT-1 relative expression quantity.
3rd, statistical procedures
Data are analyzed using SPSS 19.0.Measurement data is represented with mean value ± deviation, is examined using t, enumeration data
It is expressed as a percentage, using χ2Examine, be that difference is statistically significant with P < 0.05, and establish ROC curve, under calculated curve
Area (area under the curve, AUC) and 95% credibility interval.Selection variables are returned with Logistic, are established back
Return equation, produce one group of new variables Y.ROC curve analysis is carried out to new variables and each single index.
2nd, experimental result
1st, Luminal A types breast cancer does not shift and Bone tumour group lncRNA XLOC_004122, lncRNA SUMO1P3
With lncRNA NBAT-1 relative expression levels
In test set, lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA in each sample are determined respectively
NBAT-1 relative expression levels.Compared with the non-transfer group of Luminal A type breast cancer, Luminal A type Bone of Breast Cancer transfer groups
LncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 relative expression levels significantly raise in sample, bone
Transfer group lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 relative expression levels do not turn respectively
(1.9 ± 0.4) of shifting group relative expression levels times, (2.3 ± 0.5) times, (2.5 ± 0.4) times.
2nd, lncRNA XLOC_004122, lncRNA SUMO1P3 or lncRNA NBAT-1 relative expression levels individually use
The ROC curve analysis with the transfer of Luminal A types Bone of Breast Cancer is not shifted in diagnosis differentiation Luminal A types breast cancer
The principle of ROC curve evaluation assessment:
The Basic Evaluation index of diagnostic test has susceptibility, specificity etc., comprehensive evaluation index have Youden indexes, ROC,
AUC etc..Evaluation for diagnostic test, it is necessary first to the true group of sample to be tested is known by goldstandard.For down payment mark
Accurately fixed disease group (equivalent to the Bone of Breast Cancer transfer group in the present invention) and healthy group are (equivalent to the breast cancer in the present invention
Non- transfer group), the result detected using diagnostic test can be divided into following situation:
Positive (True Positive, TP);Diagnostic test test positive (consistent with goldstandard result);
Negative (True Negative, TN);Diagnostic test is detected as negative (consistent with goldstandard result);
False positive (False Positive, FP):Diagnostic test test positive (inconsistent with goldstandard result);
False negative (False Negative, FN):Diagnostic test is detected as negative (inconsistent with goldstandard result).
It can be represented with following table:
Susceptibility=A/ (A+C) of diagnostic test;Specificity=D/ (B+D) of diagnostic test.By susceptibility and specifically
Property can draw diagnosis sensitivity level and specific degree of the diagnostic test relative to goldstandard.High represent of susceptibility examines disease example
Break few for the number of feminine gender, rate of missed diagnosis is low;The number that healthy example is diagnosed as the positive by specific high representative is few, and misdiagnosis rate is low.
The curve that ROC curve is based on above-mentioned susceptibility and specificity is drawn out.With possible diagnosis in diagnostic test
Dividing value calculates corresponding susceptibility and specificity as diagnostic points according to above table.Then, using susceptibility as ordinate,
1- specificity is abscissa, the susceptibility of each point during each diagnostic points and specificity point is marked in coordinate diagram, connection coordinate point
Smoothed curve is obtained, the curve is ROC curve.Diagnostic points are set more much closeer, and obtained ROC curve is more smooth.
ROC curve is using each testing result as possible diagnosis dividing value, and its TG-AUC AUC size shows
The size of the diagnostic test degree of accuracy.Intrinsic degree of accuracy indexs of the area AUC as diagnostic test Authentic Assessment under ROC curve
It has been be commonly recognized that, when AUC is 0.5, i.e., without diagnostic significance;AUC represents that accuracy rate of diagnosis is relatively low at 0.5~0.7;AUC exists
When 0.7~0.9, represent that diagnostic accuracy is medium;During AUC > 0.9, represent that diagnosis has higher accuracy.
It is relative that lncRNA XLOC_004122, lncRNA SUMO1P3 or lncRNA NBAT-1 are drawn in SPSS 19.0
The diagnosis differentiation Luminal A types breast cancer that expression is individually used for does not shift the ROC with the transfer of Luminal A types Bone of Breast Cancer
Curve, AUC are respectively 0.601,0.697,0.729, have relatively low or medium accuracy.
3rd, lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 relative expression levels, which combine, examines
The structure of disconnected model and distinguish Luminal A types breast cancer for diagnosing and do not shift and Luminal A types Bone of Breast Cancer transfer
ROC curve is analyzed
With the relative of lncRNA XLOC_004122, lncRNA SUMO1P3 in test set sample and lncRNA NBAT-1
Expression (sets X as independent variable1=lncRNA SUMO1P3 relative expression levels, X2=lncRNA NBAT-1 relative expressions
Level, X3=lncRNA XLOC_004122 relative expression levels), with group, (i.e. according to goldstandard, the sample belongs to Bone tumour
Organize still non-transfer group) dependent variable is used as, to lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1
Relative expression levels' progress two during sample is shifted with Luminal A types Bone of Breast Cancer is not shifted in Luminal A types breast cancer
Metalogic returns, and obtains dualistic logistic regression equation:Y=-2.577+2.045X1+1.956X2+1.676X3;Again by each sample
LncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 relative expression levels substitute into the binary logic
Regression equation, you can the regressand value Y of each sample is obtained, using possible regressand value Y as diagnostic points, meter sensitivity and special
Property, ROC curve (as shown in Figure 1) is drawn accordingly, AUC 0.921, there is higher accuracy.According to the coordinate meter of ROC curve
Dimension mounting index=specificity+sensitivity -1 is calculated, corresponding Y value distinguishes Luminal for that can carry out diagnosis when tieing up mounting index maximum
The optimal cut-off values 0.598 (i.e. diagnostic threshold) of the non-transfer group of A type breast cancer and Bone tumour group.
4th, checking concentrates checking lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 with respect to table
The order of accuarcy with the transfer of Luminal A types Bone of Breast Cancer is not shifted up to horizontal Combining diagnosis differentiation Luminal A types breast cancer
Concentrated in checking, by each sample lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 phases
Above-mentioned regression model is substituted into expression, the regressand value Y, Y for obtaining each sample are predicted as higher than diagnostic threshold 0.598
Luminal A types Bone of Breast Cancer shifts, and the Luminal A types breast cancer that is predicted as less than diagnostic threshold 0.598 does not shift, accurately
Spend for 98.2% (108/110), as shown in Figure 2.
Embodiment 2:Luminal Type Bs Bone of Breast Cancer shifts
First, experiment sample and experimental method
1st, experiment sample
The non-transfer group of breast cancer, the test set of Luminal Type B Bone of Breast Cancer transfer groups and checking in Luminal Type Bs
Collection.
2nd, RNA extractings and qRT-PCR
Total serum IgE is extracted from serum sample using Trizol reagents (Invitrogen, Chinese Shanghai).Use NanoDrop
ND-2000 spectrophotometers (Thermo Scientific) determine the concentration and purity of total serum IgE at 260nm, and electrophoresis detection shows
Show, the RNA mass of purification well carries out subsequent operation afterwards.Total serum IgE is turned using reverse transcription reagent box (Takara, DaLian, China)
Turn to cDNA.Quantitative fluorescent PCR (Takara, DaLian, China), and application ABI Prism7000 are carried out using SYBR Green methods
Quantitative fluorescent PCR system (Agilent Technologies) carries out Data Collection.Primer is closed by winning still biological (Chinese Shanghai)
Into lncRNA XLOC_004122 primer sequences:- the CTGGCAGGAACACCGGGTACTT-3 ' of upstream 5 ', downstream 5 '-
TGACTTTTACTTAGGAGCCACTTCTTG-3’;LncRNA Linc00467 primer sequences:Upstream 5 '-
GCCTGGTTGTTCAGCACCTTCG-3 ' ,-TCGGATCGGTGCTGGTTTTGGT-3 ' of downstream 5 ';LncRNA Al049452 draw
Thing sequence:- the CAGTTAAACCCACAGGTGGTAGCATGAC-3 ' of upstream 5 ', downstream 5 '-
TAGTGGGAAAACCTAGTTTCCGACAGTT-3’;GAPDH primer sequences:- the CGCTCTCTGCTCCTCCTGTTC- of upstream 5 '
3 ' ,-ATCCGTTGACTCCGACCTTCAC-3 ' of downstream 5 '.3 average values measured are taken with 2-ΔΔCtMethod calculates lncRNA
XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 relative expression quantity.
3rd, statistical procedures
Data are analyzed using SPSS 19.0.Measurement data is represented with mean value ± deviation, is examined using t, enumeration data
It is expressed as a percentage, using χ2Examine, be that difference is statistically significant with P < 0.05, and establish ROC curve, under calculated curve
Area (area under the curve, AUC) and 95% credibility interval.Selection variables are returned with Logistic, are established back
Return equation, produce one group of new variables Y.ROC curve analysis is carried out to new variables and each single index.
2nd, experimental result
1st, Luminal Type Bs breast cancer does not shift and Bone tumour group lncRNA XLOC_004122, lncRNA
Linc00467 and lncRNA Al049452 relative expression levels
In test set, determine respectively each sample lncRNA XLOC_004122, lncRNA Linc00467 and
LncRNA Al049452 relative expression levels.Compared with the non-transfer group of Luminal Type B breast cancer, Luminal Type B breast cancer
LncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 relative expression's water in Bone tumour group sample
HUD writes up-regulation, and Bone tumour group lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 are with respect to table
Up to it is horizontal be respectively (2.2 ± 0.4) times of non-transfer group relative expression levels, (2.7 ± 0.3) times, (2.3 ± 0.3) times.
2nd, lncRNA XLOC_004122, lncRNA Linc00467 or lncRNA Al049452 relative expression levels are single
It is solely used in diagnosis differentiation Luminal Type Bs breast cancer and does not shift the ROC curve analysis shifted with Luminal Type Bs Bone of Breast Cancer
The principle of ROC curve evaluation assessment:
The Basic Evaluation index of diagnostic test has susceptibility, specificity etc., comprehensive evaluation index have Youden indexes, ROC,
AUC etc..Evaluation for diagnostic test, it is necessary first to the true group of sample to be tested is known by goldstandard.For down payment mark
Accurately fixed disease group (equivalent to the Bone of Breast Cancer transfer group in the present invention) and healthy group are (equivalent to the breast cancer in the present invention
Non- transfer group), the result detected using diagnostic test can be divided into following situation:
Positive (True Positive, TP);Diagnostic test test positive (consistent with goldstandard result);
Negative (True Negative, TN);Diagnostic test is detected as negative (consistent with goldstandard result);
False positive (False Positive, FP):Diagnostic test test positive (inconsistent with goldstandard result);
False negative (False Negative, FN):Diagnostic test is detected as negative (inconsistent with goldstandard result).
It can be represented with following table:
Susceptibility=A/ (A+C) of diagnostic test;Specificity=D/ (B+D) of diagnostic test.By susceptibility and specifically
Property can draw diagnosis sensitivity level and specific degree of the diagnostic test relative to goldstandard.High represent of susceptibility examines disease example
Break few for the number of feminine gender, rate of missed diagnosis is low;The number that healthy example is diagnosed as the positive by specific high representative is few, and misdiagnosis rate is low.
The curve that ROC curve is based on above-mentioned susceptibility and specificity is drawn out.With possible diagnosis in diagnostic test
Dividing value calculates corresponding susceptibility and specificity as diagnostic points according to above table.Then, using susceptibility as ordinate,
1- specificity is abscissa, the susceptibility of each point during each diagnostic points and specificity point is marked in coordinate diagram, connection coordinate point
Smoothed curve is obtained, the curve is ROC curve.Diagnostic points are set more much closeer, and obtained ROC curve is more smooth.
ROC curve is using each testing result as possible diagnosis dividing value, and its TG-AUC AUC size shows
The size of the diagnostic test degree of accuracy.Intrinsic degree of accuracy indexs of the area AUC as diagnostic test Authentic Assessment under ROC curve
It has been be commonly recognized that, when AUC is 0.5, i.e., without diagnostic significance;AUC represents that accuracy rate of diagnosis is relatively low at 0.5~0.7;AUC exists
When 0.7~0.9, represent that diagnostic accuracy is medium;During AUC > 0.9, represent that diagnosis has higher accuracy.
LncRNA XLOC_004122, lncRNA Linc00467 or lncRNA Al049452 are drawn in SPSS 19.0
Relative expression levels be individually used for diagnosis distinguish Luminal Type Bs breast cancer do not shift and Luminal Type Bs Bone of Breast Cancer transfer
ROC curve, AUC is respectively 0.687,0.744,0.706, has relatively low or medium accuracy.
3rd, lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 relative expression levels connection
Close the structure of diagnostic model and turn for diagnosing to distinguish Luminal Type Bs breast cancer and do not shift with Luminal Type Bs Bone of Breast Cancer
The ROC curve analysis of shifting
With lncRNA XLOC_004122, lncRNA Linc00467 in test set sample and lncRNA Al049452
Relative expression levels (set X as independent variable1=lncRNA Linc00467 relative expression levels, X2=lncRNA Al049452
Relative expression levels, X3=lncRNA XLOC_004122 relative expression levels), with group (i.e. according to the goldstandard sample category
In Bone tumour group still non-transfer group) be used as dependent variable, to lncRNA XLOC_004122, lncRNA Linc00467 and
LncRNA Al049452 do not shift relative with Luminal Type Bs Bone of Breast Cancer transfer sample in Luminal Type Bs breast cancer
Expression carries out dualistic logistic regression, obtains dualistic logistic regression equation:Y=-2.241+1.883X1+2.275X2+
1.975X3;Again by lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 phase in each sample
The dualistic logistic regression equation is substituted into expression, you can obtain the regressand value Y of each sample, make with possible regressand value Y
For diagnostic points, meter sensitivity and specificity, ROC curve (as shown in Figure 3) is drawn accordingly, AUC 0.935, is had higher
Accuracy.Dimension mounting index=specificity+sensitivity -1, corresponding Y when tieing up mounting index maximum are calculated according to the coordinate of ROC curve
It is worth for the optimal cut-off values 0.607 of the non-transfer group of diagnosis differentiation Luminal Type B breast cancer and Bone tumour group can be carried out (i.e.
Diagnostic threshold).
4th, checking lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 phases are concentrated in checking
To expression Combining diagnosis distinguish Luminal Type Bs breast cancer do not shift and Luminal Type Bs Bone of Breast Cancer transfer it is accurate
Degree
Concentrated in checking, by each sample lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA
Al049452 relative expression levels substitute into above-mentioned regression model, obtain the regressand value Y, Y of each sample higher than diagnostic threshold 0.607
The transfer of Luminal Type Bs Bone of Breast Cancer is predicted as, the Luminal Type Bs breast cancer that is predicted as less than diagnostic threshold 0.607 does not turn
Move, the degree of accuracy is 95.2% (60/63), as shown in Figure 4.
Embodiment 3:Her-2 overexpression types Bone of Breast Cancer shifts
First, experiment sample and experimental method
1st, experiment sample
The non-transfer group of breast cancer in Her-2 overexpression types, Her-2 overexpression type Bone of Breast Cancer transfer groups test set and test
Card collection.
2nd, RNA extractings and qRT-PCR
Total serum IgE is extracted from serum sample using Trizol reagents (Invitrogen, Chinese Shanghai).Use NanoDrop
ND-2000 spectrophotometers (Thermo Scientific) determine the concentration and purity of total serum IgE at 260nm, and electrophoresis detection shows
Show, the RNA mass of purification well carries out subsequent operation afterwards.Total serum IgE is turned using reverse transcription reagent box (Takara, DaLian, China)
Turn to cDNA.Quantitative fluorescent PCR (Takara, DaLian, China), and application ABI Prism7000 are carried out using SYBR Green methods
Quantitative fluorescent PCR system (Agilent Technologies) carries out Data Collection.Primer is closed by winning still biological (Chinese Shanghai)
Into lncRNA AK043773 primer sequences:- the GTGACGCCAGGGATGGCATTA-3 ' of upstream 5 ', downstream 5 '-
CAGAGCCTTGCATTGGTCAGT-3’;LncRNA EXOC7 primer sequences:Upstream 5 '-
GAGTCTGGGATCAGAGAGCAAAGG-3 ' ,-GGTACTGTAGAAAGGCCCCGTAGG-3 ' of downstream 5 ';GAPDH primer sequences:
- the CGCTCTCTGCTCCTCCTGTTC-3 ' of upstream 5 ' ,-ATCCGTTGACTCCGACCTTCAC-3 ' of downstream 5 '.Take 3 measurements
Average value is with 2-ΔΔCtMethod calculates lncRNA AK043773 and lncRNA EXOC7 relative expression quantity.
3rd, statistical procedures
Data are analyzed using SPSS 19.0.Measurement data is represented with mean value ± deviation, is examined using t, enumeration data
It is expressed as a percentage, using χ2Examine, be that difference is statistically significant with P < 0.05, and establish ROC curve, under calculated curve
Area (area under the curve, AUC) and 95% credibility interval.Selection variables are returned with Logistic, are established back
Return equation, produce one group of new variables Y.ROC curve analysis is carried out to new variables and each single index.
2nd, experimental result
1st, the non-transfer group of Her-2 overexpressions type breast cancer and Bone tumour group lncRNA AK043773 and lncRNA EXOC7
Relative expression levels
In test set, lncRNA AK043773 and lncRNA the EXOC7 relative expression levels of each sample are determined respectively.
Compared with the non-transfer group of Her-2 overexpression type breast cancer, lncRNA in Her-2 overexpression type Bone of Breast Cancer transfer group samples
AK043773 and lncRNA EXOC7 relative expression levels significantly raise, Bone tumour group lncRNA AK043773 and lncRNA
EXOC7 relative expression levels are respectively (3.3 ± 0.5) times of non-transfer group, (2.6 ± 0.3) times.
2nd, lncRNA AK043773 or lncRNA EXOC7 relative expression levels are individually used for diagnosis differentiation Her-2 and cross table
The ROC curve analysis with the transfer of Her-2 overexpression types Bone of Breast Cancer is not shifted up to type breast cancer
The principle of ROC curve evaluation assessment:
The Basic Evaluation index of diagnostic test has susceptibility, specificity etc., comprehensive evaluation index have Youden indexes, ROC,
AUC etc..Evaluation for diagnostic test, it is necessary first to the true group of sample to be tested is known by goldstandard.For down payment mark
Accurately fixed disease group (equivalent to the Bone of Breast Cancer transfer group in the present invention) and healthy group are (equivalent to the breast cancer in the present invention
Non- transfer group), the result detected using diagnostic test can be divided into following situation:
Positive (True Positive, TP);Diagnostic test test positive (consistent with goldstandard result);
Negative (True Negative, TN);Diagnostic test is detected as negative (consistent with goldstandard result);
False positive (False Positive, FP):Diagnostic test test positive (inconsistent with goldstandard result);
False negative (False Negative, FN):Diagnostic test is detected as negative (inconsistent with goldstandard result).
It can be represented with following table:
Susceptibility=A/ (A+C) of diagnostic test;Specificity=D/ (B+D) of diagnostic test.By susceptibility and specifically
Property can draw diagnosis sensitivity level and specific degree of the diagnostic test relative to goldstandard.High represent of susceptibility examines disease example
Break few for the number of feminine gender, rate of missed diagnosis is low;The number that healthy example is diagnosed as the positive by specific high representative is few, and misdiagnosis rate is low.
The curve that ROC curve is based on above-mentioned susceptibility and specificity is drawn out.With possible diagnosis in diagnostic test
Dividing value calculates corresponding susceptibility and specificity as diagnostic points according to above table.Then, using susceptibility as ordinate,
1- specificity is abscissa, the susceptibility of each point during each diagnostic points and specificity point is marked in coordinate diagram, connection coordinate point
Smoothed curve is obtained, the curve is ROC curve.Diagnostic points are set more much closeer, and obtained ROC curve is more smooth.
ROC curve is using each testing result as possible diagnosis dividing value, and its TG-AUC AUC size shows
The size of the diagnostic test degree of accuracy.Intrinsic degree of accuracy indexs of the area AUC as diagnostic test Authentic Assessment under ROC curve
It has been be commonly recognized that, when AUC is 0.5, i.e., without diagnostic significance;AUC represents that accuracy rate of diagnosis is relatively low at 0.5~0.7;AUC exists
When 0.7~0.9, represent that diagnostic accuracy is medium;During AUC > 0.9, represent that diagnosis has higher accuracy.
LncRNA AK043773 or lncRNA EXOC7 relative expression levels are drawn in SPSS 19.0 to be individually used for examining
Disconnected Her-2 overexpression type breast cancer of distinguishing does not shift the ROC curve shifted with Her-2 overexpression types Bone of Breast Cancer, and AUC is respectively
0.762nd, 0.717, there is medium accuracy.
3rd, the structure of lncRNA AK043773 and lncRNA EXOC7 relative expression levels' Combining diagnosis models and for examining
Disconnected Her-2 overexpression type breast cancer of distinguishing does not shift the ROC curve analysis shifted with Her-2 overexpression types Bone of Breast Cancer
Independent variable is used as using the relative expression levels of lncRNA AK043773 and lncRNA EXOC7 in test set sample
(set X1=lncRNA AK043773 relative expression levels, X2=lncRNA EXOC7 relative expression levels), with group (i.e. basis
The goldstandard sample belongs to Bone tumour group still non-transfer group) dependent variable is used as, to lncRNA AK043773 and lncRNA
EXOC7 does not shift relative expression's water in shifting sample with Her-2 overexpression types Bone of Breast Cancer in Her-2 overexpression type breast cancer
It is flat to carry out dualistic logistic regression, obtain dualistic logistic regression equation:Y=-2.918+2.618X1+2.115X2;Again by each sample
LncRNA AK043773 and lncRNA EXOC7 relative expression levels substitute into the dualistic logistic regression equation, you can obtain each
The regressand value Y of individual sample, using possible regressand value Y as diagnostic points, meter sensitivity and specificity, ROC curve is drawn accordingly
(as shown in Figure 5), AUC 0.939, there is higher accuracy.Further according to the coordinate of ROC curve calculate dimension mounting index=
Specificity+sensitivity -1, corresponding Y value does not turn for that can diagnose differentiation Her-2 overexpression type breast cancer when tieing up mounting index maximum
The optimal cut-off values 0.495 (i.e. diagnostic threshold) of shifting group and Bone tumour group.
4th, checking concentrates checking lncRNA AK043773 and lncRNA EXOC7 relative expression levels Combining diagnosis to distinguish
Her-2 overexpression type breast cancer does not shift the order of accuarcy with the transfer of Her-2 overexpression types Bone of Breast Cancer
Concentrated in checking, each sample lncRNA AK043773 and EXOC7 relative expression levels are substituted into above-mentioned recurrence mould
Type, the regressand value Y, Y for obtaining each sample shift higher than the Her-2 overexpression types Bone of Breast Cancer that is predicted as of diagnostic threshold 0.495, low
Do not shifted in the Her-2 overexpression type breast cancer that is predicted as of diagnostic threshold 0.495, the degree of accuracy is 91.1% (51/56), such as Fig. 6.
Embodiment 4:Triple negative breast cancer Bone tumour
First, experiment sample and experimental method
1st, experiment sample
The non-transfer group of breast cancer, the test set of three negative type breast cancers Bone tumour groups and checking collection in three negative types.
2nd, RNA extractings and qRT-PCR
Total serum IgE is extracted from serum sample using Trizol reagents (Invitrogen, Chinese Shanghai).Use NanoDrop
ND-2000 spectrophotometers (Thermo Scientific) determine the concentration and purity of total serum IgE at 260nm, and electrophoresis detection shows
Show, the RNA mass of purification well carries out subsequent operation afterwards.Total serum IgE is turned using reverse transcription reagent box (Takara, DaLian, China)
Turn to cDNA.Quantitative fluorescent PCR (Takara, DaLian, China), and application ABI Prism7000 are carried out using SYBR Green methods
Quantitative fluorescent PCR system (Agilent Technologies) carries out Data Collection.Primer is closed by winning still biological (Chinese Shanghai)
Into lncRNA Lnc01089 primer sequences:- the TCGCTGGGTTGCTCTGCTTC-3 ' of upstream 5 ', downstream 5 '-
GTCAGGAGGTCACAGTCTTAGGG-3’;LncRNA HOTAIR primer sequences:Upstream 5 '-
CGTGGAAAGATCCAAATGGGACCA-3 ' ,-AGCCTAGGAATCAGCACGAAGCAAA-3 ' of downstream 5 ';GAPDH primer sequences
Row:- the CGCTCTCTGCTCCTCCTGTTC-3 ' of upstream 5 ' ,-ATCCGTTGACTCCGACCTTCAC-3 ' of downstream 5 '.Take 3 measurements
Average value with 2-ΔΔCtMethod calculates lncRNA Lnc01089 and lncRNA HOTAIR relative expression quantity.
3rd, statistical procedures
Data are analyzed using SPSS 19.0.Measurement data is represented with mean value ± deviation, is examined using t, enumeration data
It is expressed as a percentage, using χ2Examine, be that difference is statistically significant with P < 0.05, and establish ROC curve, under calculated curve
Area (area under the curve, AUC) and 95% credibility interval.Selection variables are returned with Logistic, are established back
Return equation, produce one group of new variables Y.ROC curve analysis is carried out to new variables and each single index.
2nd, experimental result
1st, the non-transfer group of three negative type breast cancers and Bone tumour group lncRNA Lnc01089 and HOTAIR relative expression levels
In test set, lncRNA Lnc01089 and lncRNA HOTAIR relative expression's water of each sample is determined respectively
It is flat.Compared with the non-transfer group of three negative type breast cancers, in three negative type breast cancers Bone tumour group samples lncRNA Lnc01089 and
LncRNA HOTAIR relative expression levels significantly raise, and Bone tumour group lncRNA Lnc01089 and lncRNA HOTAIR are relative
Expression is respectively (3.5 ± 0.6) times of non-transfer group, (3.2 ± 0.5) times.
2nd, lncRNA Lnc01089 or lncRNA HOTAIR relative expression levels are individually used for diagnosis and distinguish three negative types
Breast cancer does not shift to be analyzed with the ROC curve of three negative type breast cancers Bone tumours
The principle of ROC curve evaluation assessment:
The Basic Evaluation index of diagnostic test has susceptibility, specificity etc., comprehensive evaluation index have Youden indexes, ROC,
AUC etc..Evaluation for diagnostic test, it is necessary first to the true group of sample to be tested is known by goldstandard.For down payment mark
Accurately fixed disease group (equivalent to the Bone of Breast Cancer transfer group in the present invention) and healthy group are (equivalent to the breast cancer in the present invention
Non- transfer group), the result detected using diagnostic test can be divided into following situation:
Positive (True Positive, TP);Diagnostic test test positive (consistent with goldstandard result);
Negative (True Negative, TN);Diagnostic test is detected as negative (consistent with goldstandard result);
False positive (False Positive, FP):Diagnostic test test positive (inconsistent with goldstandard result);
False negative (False Negative, FN):Diagnostic test is detected as negative (inconsistent with goldstandard result).
It can be represented with following table:
Susceptibility=A/ (A+C) of diagnostic test;Specificity=D/ (B+D) of diagnostic test.By susceptibility and specifically
Property can draw diagnosis sensitivity level and specific degree of the diagnostic test relative to goldstandard.High represent of susceptibility examines disease example
Break few for the number of feminine gender, rate of missed diagnosis is low;The number that healthy example is diagnosed as the positive by specific high representative is few, and misdiagnosis rate is low.
The curve that ROC curve is based on above-mentioned susceptibility and specificity is drawn out.With possible diagnosis in diagnostic test
Dividing value calculates corresponding susceptibility and specificity as diagnostic points according to above table.Then, using susceptibility as ordinate,
1- specificity is abscissa, the susceptibility of each point during each diagnostic points and specificity point is marked in coordinate diagram, connection coordinate point
Smoothed curve is obtained, the curve is ROC curve.Diagnostic points are set more much closeer, and obtained ROC curve is more smooth.
ROC curve is using each testing result as possible diagnosis dividing value, and its TG-AUC AUC size shows
The size of the diagnostic test degree of accuracy.Intrinsic degree of accuracy indexs of the area AUC as diagnostic test Authentic Assessment under ROC curve
It has been be commonly recognized that, when AUC is 0.5, i.e., without diagnostic significance;AUC represents that accuracy rate of diagnosis is relatively low at 0.5~0.7;AUC exists
When 0.7~0.9, represent that diagnostic accuracy is medium;During AUC > 0.9, represent that diagnosis has higher accuracy.
LncRNA Lnc01089 or lncRNA HOTAIR relative expression levels are drawn in SPSS 19.0 to be individually used for examining
Disconnected three negative type breast cancers of distinguishing do not shift ROC curve with three negative type breast cancers Bone tumours, AUC is respectively 0.755,
0.732, it is respectively provided with medium accuracy.
3rd, the structure of lncRNA Lnc01089 and lncRNA HOTAIR relative expression levels' Combining diagnosis models and it is used for
Diagnosis is distinguished three negative type breast cancers and not shifted and the analysis of the ROC curve of three negative type breast cancers Bone tumours
Independent variable is used as using the relative expression levels of lncRNA Lnc01089 and lncRNA HOTAIR in test set sample
(set X1=lncRNA Lnc01089 relative expression levels, X2=lncRNA HOTAIR relative expression levels), with group (i.e. root
According to goldstandard, the sample belongs to Bone tumour group still non-transfer group) dependent variable is used as, to lncRNA Lnc01089 and lncRNA
HOTAIR is not shifted in three negative type breast cancers and is carried out two with the relative expression levels in three negative type breast cancers Bone tumour samples
Metalogic returns, and obtains dualistic logistic regression equation:Y=-2.537+2.793X1+2.181X2;Again by lncRNA in each sample
Lnc01089 and lncRNA HOTAIR relative expression levels substitute into the dualistic logistic regression equation, you can obtain each sample
Regressand value Y, using possible regressand value Y as diagnostic points, meter sensitivity and specificity, draw ROC curve (such as Fig. 7 accordingly
It is shown), AUC 0.948, there is higher accuracy.Dimension mounting index=specificity is further calculated according to the coordinate of ROC curve
+ sensitivity -1, corresponding Y value distinguishes the non-transfer group of three negative type breast cancers and bone for that can carry out diagnosis when tieing up mounting index maximum
The optimal cut-off values 0.633 (i.e. diagnostic threshold) of transfer group.
4th, checking concentrates checking lncRNA Lnc01089 and lncRNA HOTAIR relative expression levels Combining diagnosis to distinguish
Three negative type breast cancers do not shift the order of accuarcy with three negative type breast cancers Bone tumours
Concentrate, each sample lncRNA Lnc01089 and lncRNA HOTAIR relative expression levels are substituted into above-mentioned in checking
Regression model, the regressand value Y, Y for obtaining each sample are predicted as three negative type breast cancers Bone tumours higher than diagnostic threshold 0.633,
Three negative type breast cancers that are predicted as less than diagnostic threshold 0.633 do not shift, and the degree of accuracy is 92.9% (52/56), such as Fig. 8.
Embodiment 5:The diagnostic kit of diagnosis indication different subtype Bone of Breast Cancer transfer
1st, Luminal A types Bone of Breast Cancer transfer diagnosis indication kit
QPCR primers including lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1:lncRNA
XLOC_004122 qPCR sense primers are as shown in Sequence NO.1, and qPCR anti-sense primers are as shown in Sequence NO.2;
LncRNA SUMO1P3 qPCR sense primers are as shown in Sequence NO.3, qPCR anti-sense primers such as Sequence NO.4 institutes
Show;LncRNA NBAT-1 qPCR sense primers are as shown in Sequence NO.5, qPCR anti-sense primers such as Sequence NO.6
It is shown.Also include internal reference GAPDH qPCR primers:Internal reference GAPDH qPCR sense primers as shown in Sequence NO.19,
QPCR anti-sense primers are as shown in Sequence NO.20.
Also include other qRT-PCR reagents such as the enzyme needed for qRT-PCR.
2nd, Luminal Type Bs Bone of Breast Cancer transfer diagnosis indication kit
QPCR primers including lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452:
LncRNA XLOC_004122 qPCR sense primers are as shown in Sequence NO.1, qPCR anti-sense primers such as Sequence
Shown in NO.2;LncRNA Linc00467 qPCR sense primers are as shown in Sequence NO.7, and qPCR anti-sense primers are such as
Shown in Sequence NO.8;As shown in Sequence NO.9, qPCR draws in downstream lncRNA Al049452 qPCR sense primers
Thing is as shown in Sequence NO.10.Also include internal reference GAPDH qPCR primers:Internal reference GAPDH qPCR sense primers are such as
Shown in Sequence NO.19, qPCR anti-sense primers are as shown in Sequence NO.20.
Also include other qRT-PCR reagents such as the enzyme needed for qRT-PCR.
3rd, Her-2 overexpressions type Bone of Breast Cancer transfer diagnosis indication kit
Include lncRNA AK043773 and lncRNA EXOC7 qPCR primers:LncRNA AK043773 qPCR upstreams
Primer is as shown in Sequence NO.11, and qPCR anti-sense primers are as shown in Sequence NO.12;LncRNA EXOC7 qPCR
Sense primer is as shown in Sequence NO.13, and qPCR anti-sense primers are as shown in Sequence NO.14.Also include internal reference GAPDH
QPCR primers:Internal reference GAPDH qPCR sense primers are as shown in Sequence NO.19, qPCR anti-sense primers such as Sequence
Shown in NO.20.
Also include other qRT-PCR reagents such as the enzyme needed for qRT-PCR.
4th, three negative type breast cancers Bone tumours diagnosis indication kit
Include lncRNA Lnc01089 and lncRNA HOTAIR qPCR primers:On lncRNA Lnc01089 qPCR
Primer is swum as shown in Sequence NO.15, qPCR anti-sense primers are as shown in Sequence NO.16;LncRNA HOTAIR's
QPCR sense primers are as shown in Sequence NO.17, and qPCR anti-sense primers are as shown in Sequence NO.18.Also include internal reference
GAPDH qPCR primers:Internal reference GAPDH qPCR sense primers are as shown in Sequence NO.19, and qPCR anti-sense primers are such as
Shown in Sequence NO.20.
Also include other qRT-PCR reagents such as the enzyme needed for qRT-PCR.
In summary, it is a discovery of the invention that serum lncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA
NBAT-1 can combine for diagnose indication Luminal A types breast cancer whether Bone tumour, concentrate diagnosis indication in individual authentication
Rate of accuracy reached more than 90%;Serum lncRNA XLOC_004122, lncRNA Linc00467 and lncRNA Al049452 can be with
Combine for diagnose indication Luminal Type Bs breast cancer whether Bone tumour, concentrate diagnosis indication rate of accuracy reached in individual authentication
More than 90%;Serum lncRNA AK043773 and lncRNA EXOC7 can combine for diagnosing indication Her-2 overexpressions type breast
Gland cancer whether Bone tumour, individual authentication concentrate diagnosis indication rate of accuracy reached more than 90%;LncRNA Lnc01089 and lncRNA
HOTAIR can combine for diagnose three negative type breast cancers of indication whether Bone tumour, concentrate diagnosis indication accurate in individual authentication
Rate is up to more than 90%.Diagnosing the transfer of indication different molecular hypotype Bone of Breast Cancer using above-mentioned serum lncRNA, not only the degree of accuracy is high,
And testing cost is low, non-invasi, convenient and swift, patient suffering and burden are greatly reduced.
The effect of above-described embodiment is the specific essentiality content for introducing the present invention, but those skilled in the art should know
Road, protection scope of the present invention should not be confined to the specific embodiment.
Sequence table
<110>Li Yijian
<120>LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer transfering reagent boxes
<160> 20
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
ctggcaggaa caccgggtac tt 22
<210> 2
<211> 27
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
tgacttttac ttaggagcca cttcttg 27
<210> 3
<211> 23
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
ctggaactgg gaatggagga aga 23
<210> 4
<211> 23
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
gattgagaaa ggattgaggg aaa 23
<210> 5
<211> 23
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
ctgggaaagc ctgtgctctt gga 23
<210> 6
<211> 23
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
gcttcacagt gctgctcaat cgt 23
<210> 7
<211> 22
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
gcctggttgt tcagcacctt cg 22
<210> 8
<211> 22
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 8
tcggatcggt gctggttttg gt 22
<210> 9
<211> 28
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 9
cagttaaacc cacaggtggt agcatgac 28
<210> 10
<211> 28
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 10
tagtgggaaa acctagtttc cgacagtt 28
<210> 11
<211> 21
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 11
gtgacgccag ggatggcatt a 21
<210> 12
<211> 21
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 12
cagagccttg cattggtcag t 21
<210> 13
<211> 24
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 13
gagtctggga tcagagagca aagg 24
<210> 14
<211> 24
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 14
ggtactgtag aaaggccccg tagg 24
<210> 15
<211> 20
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 15
tcgctgggtt gctctgcttc 20
<210> 16
<211> 23
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 16
gtcaggaggt cacagtctta ggg 23
<210> 17
<211> 24
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 17
cgtggaaaga tccaaatggg acca 24
<210> 18
<211> 25
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 18
agcctaggaa tcagcacgaa gcaaa 25
<210> 19
<211> 21
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 19
cgctctctgc tcctcctgtt c 21
<210> 20
<211> 22
<212> DNA/RNA
<213>Artificial sequence (Artificial Sequence)
<400> 20
atccgttgac tccgaccttc ac 22
Claims (10)
- A kind of 1. lncRNA diagnosis compositions, it is characterised in that:By lncRNA XLOC_004122, lncRNA SUMO1P3 and LncRNA NBAT-1 are formed.
- 2. the diagnosis composition described in claim 1 is preparing the diagnosis examination of diagnosis indication Luminal A types Bone of Breast Cancer transfer Application in terms of agent box.
- A kind of 3. diagnostic kit for being used to diagnose the Bone of Breast Cancer transfer of indication LuminalA types, it is characterised in that:Including LncRNA XLOC_004122, lncRNA SUMO1P3 and lncRNA NBAT-1 qPCR primers.
- 4. diagnostic kit according to claim 3, it is characterised in that:Draw lncRNA XLOC_004122 qPCR upstreams Thing is as shown in Sequence NO.1, and qPCR anti-sense primers are as shown in Sequence NO.2.
- 5. diagnostic kit according to claim 3, it is characterised in that:LncRNA SUMO1P3 qPCR sense primers are such as Shown in Sequence NO.3, qPCR anti-sense primers are as shown in Sequence NO.4.
- 6. diagnostic kit according to claim 3, it is characterised in that:LncRNANBAT-1 qPCR sense primers are such as Shown in Sequence NO.5, qPCR anti-sense primers are as shown in Sequence NO.6.
- 7. diagnostic kit according to claim 3, it is characterised in that:Also include internal reference GAPDH qPCR primers.
- 8. diagnostic kit according to claim 7, it is characterised in that:The qPCR sense primers of the internal reference GAPDH are such as Shown in Sequence NO.19, qPCR anti-sense primers are as shown in Sequence NO.20.
- 9. according to any described diagnostic kits of claim 3-8, it is characterised in that:Also include the enzyme needed for qRT-PCR.
- A kind of 10. method for diagnosing the Bone of Breast Cancer transfer of indication LuminalA types, it is characterised in that comprise the following steps:Step S1, LuminalA type patient with breast cancer's limosis vein bloods are gathered, serum is centrifuged out after natural coagulation;Step S2, serum total serum IgE is extracted, lncRNA XLOC_004122, lncRNA in total serum IgE are determined with qRT-PCR methods SUMO1P3 and lncRNA NBAT-1 use X successively relative to internal reference GAPDH relative expression levels3、X1、X2Represent;Step S3, by X3、X1、X2Value substitutes into dualistic logistic regression equation Y=-2.577+2.045X1+1.956X2+1.676X3 To Y value, Y value is more than 0.598 and indicates that Bone tumour occurs for the patient with breast cancer, and Bone tumour does not occur less than 0.598 indication.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201711142657.1A CN107746887B (en) | 2017-11-17 | 2017-11-17 | LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer metastatic gene diagnostic kits |
PCT/CN2018/072269 WO2019095541A1 (en) | 2017-11-17 | 2018-01-11 | Composition and method for diagnosing and predicting breast cancer bone metastases |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201711142657.1A CN107746887B (en) | 2017-11-17 | 2017-11-17 | LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer metastatic gene diagnostic kits |
Publications (2)
Publication Number | Publication Date |
---|---|
CN107746887A true CN107746887A (en) | 2018-03-02 |
CN107746887B CN107746887B (en) | 2018-08-28 |
Family
ID=61252228
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201711142657.1A Expired - Fee Related CN107746887B (en) | 2017-11-17 | 2017-11-17 | LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer metastatic gene diagnostic kits |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107746887B (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
CN111214482A (en) * | 2020-02-21 | 2020-06-02 | 东莞市第八人民医院(东莞市儿童医院) | Application of linc00467 gene-targeted siRNA in drug resistance of leukemia |
-
2017
- 2017-11-17 CN CN201711142657.1A patent/CN107746887B/en not_active Expired - Fee Related
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
CN111214482A (en) * | 2020-02-21 | 2020-06-02 | 东莞市第八人民医院(东莞市儿童医院) | Application of linc00467 gene-targeted siRNA in drug resistance of leukemia |
Also Published As
Publication number | Publication date |
---|---|
CN107746887B (en) | 2018-08-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105603101B (en) | Detect application of the system of 8 miRNA expression quantity in diagnosis or auxiliary diagnosis of hepatoma product is prepared | |
CN110484624B (en) | Gastric cancer biomarker based on peripheral blood and detection method and application thereof | |
CN108841962A (en) | A kind of non-small cell lung cancer detection kit and its application | |
CN102534008B (en) | SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of noncardia cancer and application thereof | |
Qiu et al. | Differential expression profiling of circulation microRNAs in PTC patients with non-131I and 131I-avid lungs metastases: a pilot study | |
CN107881239B (en) | miRNA marker related to colorectal cancer metastasis in plasma and application thereof | |
CN102534009B (en) | SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of primary lung cancer and application thereof | |
CN108531586A (en) | A kind of relevant cycle miRNA marker and its application on X chromosome of and Computer-aided Diagnosis of Breast Cancer | |
CN107746887B (en) | LncRNA compositions and the purposes for preparing diagnosis indication Luminal A type Bone of Breast Cancer metastatic gene diagnostic kits | |
CN107916291B (en) | It is a kind of for diagnose indication three negative type breast cancers Bone tumours gene diagnosis kit | |
CN104818322B (en) | MiRNA and Cyfra21 1 combine the application in detection non-small cell lung cancer | |
CN102925444A (en) | Serum micro ribonucleic acid (miRNA) biomarker of bladder cancer and detection method of expression quantity thereof | |
CN107746888B (en) | A kind of gene diagnosis kit shifted for diagnosing indication Her-2 overexpression type Bone of Breast Cancer | |
CN110257514A (en) | A kind of new cancer of the esophagus blood miRNA marker and its application | |
CN106636440B (en) | Blood plasma microRNAs is used to prepare the purposes of the diagnostic reagent of patients with lung adenocarcinoma in sieving and diagnosis male population | |
CN107699619B (en) | The purposes of lncRNA composition and preparation diagnosis indication Luminal Type B Bone of Breast Cancer metastatic gene diagnostic kit | |
CN107904309B (en) | It is a kind of for diagnose indication three negative type breast cancers Bone tumours gene diagnosis kit | |
Bustos et al. | Diagnostic miRNA signatures in paired tumor, plasma, and urine specimens from renal cell carcinoma patients | |
CN106939354A (en) | MiRNA 4530 as pulmonary cancer diagnosis mark application | |
WO2019095541A1 (en) | Composition and method for diagnosing and predicting breast cancer bone metastases | |
CN106636450B (en) | It is a kind of for diagnosing the non-invasive marker object and kit of Lung Squamous Carcinoma Patients in non-smoking or slight smoking population | |
CN107699620B (en) | A kind of gene diagnosis kit for diagnosing indication Luminal Type B Bone of Breast Cancer transfer | |
CN107881235B (en) | A kind of gene diagnosis kit shifted for diagnosing indication Luminal A type Bone of Breast Cancer | |
CN107858430B (en) | A kind of gene diagnosis kit shifted for diagnosing indication Her-2 overexpression type Bone of Breast Cancer | |
CN104878012B (en) | Applications of the 5p of Hsa miR 3200 in preparing early screening or diagnosing Brachyury positive tumors reagent or kit |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
TA01 | Transfer of patent application right |
Effective date of registration: 20180626 Address after: 242300 Ningguo city science and technology innovation service center, outer ring road, Ningguo, Xuancheng, Anhui Applicant after: Anhui Yuntai Biological Technology Co., Ltd. Address before: 226019 Nantong University, Nantong City, Jiangsu, No. 9, Nantong University Applicant before: Li Yijian |
|
TA01 | Transfer of patent application right | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180828 Termination date: 20181117 |
|
CF01 | Termination of patent right due to non-payment of annual fee |