CN108486249A - A kind of Polymorphism marker and its detection kit for prostate cancer therapy assessment - Google Patents
A kind of Polymorphism marker and its detection kit for prostate cancer therapy assessment Download PDFInfo
- Publication number
- CN108486249A CN108486249A CN201810287522.2A CN201810287522A CN108486249A CN 108486249 A CN108486249 A CN 108486249A CN 201810287522 A CN201810287522 A CN 201810287522A CN 108486249 A CN108486249 A CN 108486249A
- Authority
- CN
- China
- Prior art keywords
- artificial sequence
- mirna
- prostate cancer
- expression quantity
- genes
- 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.)
- Withdrawn
Links
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
- C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
-
- 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
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
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)
Abstract
The present invention provides the molecular marked compound combinations that can be used for carrying out drug resistance in prostate cancer chemotherapy process Pre-Evaluation, the molecular marked compound is made of 2 prostate cancer sensitive genes (P53 genes, GSTP1 genes) and 1 miRNA (miRNA 155), can be applied to the assessment to chemotherapy resistance；Meanwhile the present invention also provides the primer sets and kit applied to assessment；The present invention also provides the application of above-mentioned primer sets and kit in the joint discriminant equation of molecular labeling expression quantity.By assessing the relative expression quantity of said gene, preferably drug resistance present in docetaxel and prednisone combination therapy prostate cancer can be assessed.
Description
Technical field
It is the invention belongs to prostate cancer therapy and diagnostic techniques field, more particularly to a kind of for being imitated to prostate cancer chemotherapy
The gene association marker of fruit and drug resistance assessment, and the detection kit that is prepared by the joint marker.
Background technology
In recent years, China's prostate-cancer incidence significantly increases.Difference is constituted by stages with western countries patients with prostate cancer,
It has been just late period transfer patient when the patients with prostate cancer first visit in China nearly 70%.Although most patients are to endocrine therapy
Sensitivity, but nearly all patient can be to Hormone refractory prostate cancer (HRPC), patient survival after treating 18 months
It is usually no more than 2 years.Two results of study of TAX327 in 2004 and SWOC-9916 show the chemotherapy based on docetaxel compared with
Mitoxantrone can significantly extend the overall survival phase of HRPC patient, establish its ground as HRPC first-line treatment drugs at one stroke
Position.But clinically still there is patient the bad problem of Taxotere therapeutic effect occur, therefore, advance pair with more west he
Plug treatment drug resistance correlation circumstance carries out assessment and is of great significance.
MiRNA is highly conserved, endogenous non-coding the microRNA of one kind proved recently, in post-transcriptional level
Adjust the expression of gene.Recent studies have shown that miRNA participates in various adjusting accesses, including growth and development, cell increase
It grows and a series of important life adjustment processes such as apoptosis, signal transduction.It has been found that the abnormal expression of miRNA may lead to the mankind
A variety of diseases, such as cancer, autoimmune disease and kidney trouble.Researches show that miRNA to participate in regulation and control human body cell carefully
The processes such as born of the same parents' metabolism, increment, differentiation, apoptosis, immune；Its unconventionality expression has played key work in the occurrence and development of disease
With.Researches show that miRNA can quickly be released into blood in pathological development from tissue, in serum, blood plasma, saliva and urine,
These extracellular miRNA are related with the different pathological status of tumour.The nails such as Weber have detected 12 kinds and come from different phase pregnancy woman
MiRNA in the body fluid and urine specimen of female and different urothelial cell tumor patients, using quantitative PCR to these body fluid
In miRNA do Broad Spectrum Analysis of Infinitesimal.As a result show that miRNA exists in all body fluid, in different body fluid
Composition ratio is significantly different, some high-caliber miRNA express identical in a variety of body fluid, and some miRNA are in particular volume
It is enriched in liquid, it was further observed that the urine specimen of different physiological status has unique miRNA patterns.Prompt uses specific miR-NA
Level can be used as detecting and monitoring different pathological physiological status.
Contain a large amount of biological informations and the advantage with noninvasive collection, sample disposal relative ease, detection in urine
The tumor markers of RNA classes can be also used for diagnosing the entity tumor of other urinary systems, and this kind of cycle miRNA in urine
Unique expression pattern it is related to specific disease, the potential of especially early stage, high specific and sensibility can be used for
Neoplasm staging and early prediction draw.Bryant etc. uses 78 prostates of the qRT-PCR chip analysis containing 742 miRNA
Cancer patient and 28 normal healthy controls serum specimens, it is determined that the miRNA of 12 expression differences.And then detect 135 parts of Healthy Peoples
With 5 selected miRNA in the cancer Urine in Patients sample of forefront, miR-107 and miR-574-3p water in tumor patient urine is found
It is flat to be significantly higher than normal healthy controls, it is diagnosable go out patients with prostate cancer, and accuracy is higher than detection urine prostate specific
Antigen (PSA).
Docetaxel treatment HRPC drug-resistant performances are judged to there is no research at present using miRNA.
Invention content
The object of the present invention is to provide can be used for carrying out Pre-Evaluation to drug resistance in prostate cancer chemotherapy process dividing
Sub- marker combination, which is by 2 prostate cancer sensitive genes (P53 genes, GSTP1 genes) and 1 miRNA
(miRNA-155) it is formed, can be applied to the assessment to chemotherapy resistance；Meanwhile the present invention also provides applied to assessment
Primer sets and kit；The present invention also provides above-mentioned primer sets and kit in the joint differentiation side of molecular labeling expression quantity
Journey（Joint marker）In application.
The first aspect of the invention：
A kind of combined mark object for Hormone refractory prostate cancer chemotherapy resistance recruitment evaluation, include P53 genes,
GSTP1 genes and miRNA-155 and reference gene U6 and GAPDH.
The chemotherapy refers to docetaxel and prednisone combination therapy.
The second aspect of the invention：
A kind of reagent for being detected to above-mentioned combined mark object.
The reagent is the reagent for being detected to the expression quantity for combining marker.
Include in the reagent：
Primer pair for expanding P53 genes, nucleotide sequence is as shown in SEQ ID NO. 1~2；
Primer pair for expanding GSTP1 genes, nucleotide sequence is as shown in SEQ ID NO. 5~6；
For the primer of miRNA-155 reverse transcriptions, nucleotide sequence is as shown in SEQ ID NO. 34；
Primer pair for expanding miRNA-155 reverse transcriptions cDNA, nucleotide sequence is as shown in SEQ ID NO. 35~36；
Primer pair for expanding reference gene U6, nucleotide sequence is as shown in SEQ ID NO. 37~38；
Primer pair for expanding reference gene GAPDH, nucleotide sequence is as shown in SEQ ID NO. 39~40.
It include the kit of mentioned reagent.
Application of the above-mentioned reagent in preparing Hormone refractory prostate cancer chemotherapy resistance recruitment evaluation kit.
The application refers to whether assessing the value of following formula F more than 3.2：
F1=((- Ln (expression quantity of the miR-155 relative to U6))-0.5+ 0.71) × (Ln (expression of the TP53 relative to GAPDH
Amount)+0.24) -0.52 × Ln (expression quantity of the GSTP1 relative to GAPDH)；
When the value of F is more than 3.2, it is believed that there are drug resistance risks.
Advantageous effect
The present invention provides the molecular marked compounds that can be used for carrying out drug resistance in prostate cancer chemotherapy process Pre-Evaluation to combine,
The molecular marked compound is by 2 prostate cancer sensitive genes (P53 genes, GSTP1 genes) and 1 miRNA (miRNA-155) institute
Composition, can be applied to the assessment to chemotherapy resistance；Meanwhile the present invention also provides the primer sets and reagent applied to assessment
Box；The present invention also provides the application of above-mentioned primer sets and kit in the joint discriminant equation of molecular labeling expression quantity.It is logical
The relative expression quantity for crossing assessment said gene, can be preferably to existing in docetaxel and prednisone combination therapy prostate cancer
Drug resistance assessed.
Description of the drawings
Fig. 1 is the ROC figures of specificity and sensibility that formula 1 carries out drug resistance judgement as combined mark；
Fig. 2 is the ROC figures of specificity and sensibility that formula 2 carries out drug resistance judgement as combined mark；
Fig. 3 is the ROC figures of specificity and sensibility that formula 3 carries out drug resistance judgement as combined mark.
Specific implementation mode
Below by specific implementation mode combination attached drawing, invention is further described in detail.But those skilled in the art
It will be understood that the following example is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.It is not specified in embodiment
Particular technique or condition person, according to technology described in document in the art or condition (such as with reference to J. Pehanorm Brookers
Deng write, what Huang Peitang etc. was translated《Molecular Cloning:A Laboratory guide》, the third edition, Science Press) or according to product description into
Row.Reagents or instruments used without specified manufacturer, being can be with conventional products that are commercially available.
Embodiment 1
We selected following 3 may be relevant with tumor invasion with the relevant sensitive gene of prostate cancer and 10
MiR-96 gene analyzes itself and the drug resistant correlation of prostate cancer.Gene is as follows：
The expression quantity of said gene and miRNA are detected using quantitative fluorescent PCR, first by extracting RNA in arena,
It is transcribed into after cDNA again, carries out qPCR analyses；The reference gene used is opposite by gene and miRNA respectively for U6 and GAPDH
It is compared in their expression quantity, statistics calculates the relative expression quantity of gene and miRNA.
Wherein, primer pair is as follows used by being expanded for the cDNA of above-mentioned 3 genes：
Wherein, the process for reverse transcription being carried out to miRNA uses ring stem method, the reverse transcription primer used and is expanded to cDNA
The primer used is as follows：
Wherein, as follows for the amplimer sequence of reference gene U6 and GAPDH：
Extraction, reverse transcription and the quantitative fluorescent PCR of RNA in 1 urine of embodiment
L, 20~30ml of fore-stream urine after massage of prostate is taken, is immediately placed in ice water cooling.Under 4 DEG C, 3000r/min from
Heart 10min collects arena, and the cold D-hanks liquid of equivalent is added to wash 2 times.
2, arena is collected into 1.5ml Eppendorf pipes, that is, adds 500 μ of denaturing liquid containing acid guanidinium isothiocyanate
L mixings set -70 DEG C of preservations.
3, illustrate to be dried in vacuo after extracting total serum IgE according to classical total serum IgE extraction agent box, with 15 μ l DEPC aqueous solutions.
4, it takes 2 μ lRNA solution to add 98 μ l water, absorbance is measured on 800 type UV detectors of DU after mixing
A260/A280And rna content, 1% denaturing formaldehyde gel electroresis appraisal of RNA sample integrality.
5, reverse transcription：
Purpose dependency basis is carried out to the RNA samples of extraction using Promega GoScrip Reverse Transcriptase kits and reverse transcriptase primer
Cause, miRNA and reference gene carry out reverse transcription（Wherein, random primer reverse transcription is used for gene and reference gene, for
MiRNA carries out reverse transcription using above-mentioned miRNA specificity loop-stem structure (Stem-loops) reverse transcriptase primer）, concrete operations
It is referred to specification, whole operation process carries out on ice.
It is as follows：
(1) Reverse transcription mix (RT Master Mix) is prepared according to required number of samples, RT in each sample reaction system
Mix amounts to 6 μ L, as follows：
(2) mixing that turns upside down simultaneously centrifuges, and RT Master Mix are only sub-packed in 200 μ L without in enzyme EP pipes according to every sample 6.
(3) after centrifuging often pipe be added needed for sample each 4 μ L of RNA templates, total reverse transcription reaction system is l0 μ L, after mixing
Centrifugation.
(4) reaction condition of reverse transcription is set：25 DEG C of 1 hour → 70 DEG C of 5min → 42 DEG C 15min.After the completion of reverse transcription
The cDNA of synthesis is immediately placed on cooled on ice, is saved backup in -20 DEG C of refrigerators.
6, quantitative fluorescent PCR
Real-time fluorescence quantitative PCR reaction system：100 μ l diluted cDNA, 1ml qPCR Master Mix (2 ×), 900 μ l
ddH2O is being mixed into the mixed liquor that total volume is 2ml, adds in the corresponding each holes 20 μ l to PCR Array, PCR amplifications:①
PCR reaction systems are heated to 95 DEG C of progress pre-degenerations, 5min is reacted, is 2. cooled to 60 DEG C after reaction of degeneration (RD) 15s at 95 DEG C
Annealing reaction 32s, and be a cycle with 72 DEG C of extension 32s, so amplification 40 recycles, and finally establishes melting curve simultaneously
It identifies product specificities, and records Cq values；Wherein, the composition of qPCR Master Mix is Hot-start Taq DNA
polymerase、dNTP Mixture、MgCl2, SYBR Green, forward primer and reverse primer；Wherein forward and reverse primer is total
Amount be respectively be 2.5 μM respectively, volume be 2 μ l.
The drafting of standard curve：
It will be from Human Prostate Cancer Cells LNCaP cells（The Shanghai bio tech ltd Yan Yu is bought）The miRNA of middle extraction, it is inverse
It is transcribed into cDNA, then 10 times of gradient dilutions are at 5 concentration, and as standard items, RT-PCR amplifications are carried out together with sample to be tested,
Make standard curve.
7, result judges
(1) copy number takes logarithm as abscissa using 10 the bottom of for, and Cq values are mapped for ordinate, draw standard curve, calculate slope
S, then according to formula E=10（-1/S）- 1, calculate amplification efficiency E；
(2) sample and check and correction pattern detection hole are chosen, sample threshold Cq is obtained using the accompanying software of real-time fluorescence quantitative PCR instrument
(T) and sample threshold Cq (C) is proofreaded, according to formula Q=(E+1)-△Cq, △ Cq=[Cq (T)-Cq (C)], obtain gene correction rise
Beginning copy number Q；By the Q values of each gene and miRNA compared with the Q values of reference gene, each gene and the phase of miRNA can be obtained
To expression quantity, wherein the expression quantity of each gene is calculated relative to the expression quantity of GAPDH, the expression quantity of miRNA is opposite
It is calculated in the expression quantity of U6.
Sample information
HRPC patient 29 is accepted in selection for medical treatment, and at 61~78 years old age, 69.1 ± 10.3 years old average, all patients demonstrate,prove through pathological examination
Actually HRPC, wherein 21 patients once row bilateral orchidectomy castration, 8 once row medical castration treatments, all patients
A kind of antiandrogen drug was at least used to treat.Before chemotherapy serum PSA (PSA) it is horizontal for 39.1~
482.1 μ g/L, average 239.1 μ g/L, through sb.'s illness took a turn for the worse during endocrine therapy, (intermittent phase at least 2w) 3 times continuous is measured
Blood PSA levels increase, serum testosterone up to castration it is horizontal (<1.7nmol/L), stop anti-androgen therapy and two wires antiandrogen
After drug therapy, blood PSA values, which do not decline still, even to be risen.Select healthy person 10 as a contrast.
The therapy of HRPC patient：Docetaxel (trade name Austria name profit, Jiangsu Aosaikang Pharmaceutical Co., Ltd.'s life
Production) 75mg/m2, the 1st day 1~1.5h of intravenous drip, 1d (d0) patient's oral dexamethasone before chemotherapy, 16mg/d continues at least
3d, to prevent allergic reaction and fluid retention.Prednisone 5mg takes orally, 2 times/d, and it was 1 week that 21d is continuously taken from d1~d21
Phase.The 1st day conventional Shu Outing that gives of chemotherapy prevents vomiting reaction, as leucocyte (WBC) ＜ 2.0 × 109Take the circumstances into consideration to give grain when/L
Colony-stimulating factor supportive treatment gives imodium processing if diarrhea such as occurs in therapeutic process.It is cold that cold food is avoided during medication
Drink avoids the cold object of contact.
Evaluation curative effect and toxic side effect after all patients carry out to the chemotherapy in 5 periods.Chemotherapy promoting circulation of blood is conventional, biochemical
Routine, PSA, electrocardiogram, C-XF, Whole body bone scan, privileged site CT or MRI inspection etc..PSA is as therapeutic response
Index is widely used in the world, declines >=50% with blood-serum P SA, and can maintain one week be effective；PSA still rises
It is a height of invalid.
On inspection, it is considered as that treatment is effective to have 17 after 5 periods of chemotherapy in patient, effective percentage 60.7%.
Statistical comparison of the average expression amount of each molecular labeling between patient and healthy person（It is derived from right logarithm Ln）
*, ※ is relative to healthy person P<0.05；
# is relative to responder P<0.05.
As can be seen from the above table, for TP53 and GSTP1, they are in docetaxel treatment effectively with treatment nonresponder's
Expression quantity for healthy person, all lower respectively, prompts their expression quantity and the close phase of prostate cancer by apparent upper reconcile
It closes；Also, the expression quantity between responder and nonresponder is there is also conspicuousness difference, prompts to be possible to them to become and examine
The label of disconnected docetaxel chemotherapy resistance；And for Bcl2 genes, in the expression quantity that treatment is effective and nonresponder is with health
There are significant differences, and it is a vital signs of diagnosis of prostate cancer to prompt the variation that Bcl2 genes are；Docetaxel is one
The semi-synthetic paclitaxel derivatives of kind, pharmaceutical research show that it can be by reinforcing tubulin polymerization effect and inhibiting micro-pipe solution
Poly- effect, tumoricidal mitosis, while Bcl2 phosphorylations are may additionally facilitate, make Bcl2/Bax heterodimer depolymerization,
And then inducing apoptosis of tumour cell, for treatment responder, although also having Bcl2 amounts overexpression, how western use is
He matches bad, prompts that result in treatment invalid there may be other factors.
To sum up analyze, filtered out treatment effectively and it is invalid between there are TP53, GSTP1 of significant difference,
MiR-221, miR-155 carry out combinatorial formula evaluating and screening, are lowered according to being reconciled on expression quantity, bent by the ROC of SPSS softwares
Line is fitted, and has been gone out following 3 groups of combinatorial formulas：
；Think drug resistance when F1 is more than 3.2；（Formula
1）
；Think drug resistance when F2 is more than 1.2（Formula
2）
；Think drug resistance when F3 is more than 1.1（Formula 3）
Wherein：
A=Ln (expression quantity of the TP53 relative to GAPDH)；
B=Ln (expression quantity of the GSTP1 relative to GAPDH)；
C=Ln (expression quantity of the miR-221 relative to U6)；
D=Ln (expression quantity of the miR-155 relative to U6)；
The sensibility and specificity counted respectively by three combinatorial formulas is as follows, and ROC schemes respectively such as the institutes of Fig. 1~3
Show：
As can be seen from the table, after adding the modifying factor containing miR-155 expression quantity to the expression quantity of TP53 in formula 1,
The sensibility and specificity of diagnosis is significantly enhanced, and better than the diagnostic formulation using miR-221 expression quantity.
It in summary it can be seen, can be used for treating prostate to docetaxel using joint molecular labeling provided by the invention
The drug resistant pre- judgement of cancer.
Sequence table
<110>Mao Qiangping
<120>A kind of Polymorphism marker and its detection kit for prostate cancer therapy assessment
<130>Nothing
<150> 2017102156592
<151> 2017-04-04
<160> 40
<170> SIPOSequenceListing 1.0
<210> 1
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
tagttgtagt ggatggtggt acagt 25
<210> 2
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
actgtaccac catccactac aacta 25
<210> 3
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
tcctttcgga tctttatttc atgag 25
<210> 4
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
ctcatgaaat aaagatccga aagga 25
<210> 5
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
cagctctggc cctgatctgc cagca 25
<210> 6
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
tgctggcaga tcagggccag agctg 25
<210> 7
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgaccc cctctg 56
<210> 8
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 8
gaccccctct ggtcaaccag tcaca 25
<210> 9
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 9
tgtgactggt tgaccagagg gggtc 25
<210> 10
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 10
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacaa cccatg 56
<210> 11
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 11
gacaacccat ggaattcagt tctca 25
<210> 12
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 12
tgagaactga attccatggg ttgtc 25
<210> 13
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 13
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacac ccaccg 56
<210> 14
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 14
acacccaccg acaacaatga atgtt 25
<210> 15
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 15
aacattcatt gttgtcggtg ggtgt 25
<210> 16
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 16
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacca gctgct 56
<210> 17
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 17
accagctgct tttgggattc cgttg 25
<210> 18
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 18
caacggaatc ccaaaagcag ctggt 25
<210> 19
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 19
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacaa atctac 56
<210> 20
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 20
gacaaatcta cattgtatgc caggt 25
<210> 21
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 21
acctggcata caatgtagat ttgtc 25
<210> 22
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 22
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacag gatcta 56
<210> 23
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 23
gacaggatct acactggcta ctgag 25
<210> 24
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 24
ctcagtagcc agtgtagatc ctgtc 25
<210> 25
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 25
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacaa ctcagc 56
<210> 26
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 26
gacaactcag cttgtcaaat acacg 25
<210> 27
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 27
cgtgtatttg acaagctgag ttgtc 25
<210> 28
<211> 56
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 28
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacct gaacac 56
<210> 29
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 29
gacctgaaca ccaaaagaaa tcagt 25
<210> 30
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 30
actgatttct tttggtgttc aggtc 25
<210> 31
<211> 55
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 31
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacga acacc 55
<210> 32
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 32
gacgaacacc aggagaaatc ggtca 25
<210> 33
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 33
tgaccgattt ctcctggtgt tcgtc 25
<210> 34
<211> 54
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 34
gtcgtatcca gtgcgtgtcg tggagtcggc aattgcactg gatacgacac ccct 54
<210> 35
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 35
acacccctat cacgattagc attaa 25
<210> 36
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 36
ttaatgctaa tcgtgatagg ggtgt 25
<210> 37
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 37
catatacacg acggcttcgc tcgtg 25
<210> 38
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 38
aaatatggaa cgcttcacga att 23
<210> 39
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 39
taaaacctcc ctagagcgag g 21
<210> 40
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 40
ggagcgagat ccctccaaaa t 21
Claims (8)
1. a kind of combined mark object for Hormone refractory prostate cancer chemotherapy resistance recruitment evaluation, which is characterized in that including
There are P53 genes, GSTP1 genes and miRNA-155 and reference gene U6 and GAPDH.
2. the combined mark object according to claim 1 for Hormone refractory prostate cancer chemotherapy resistance recruitment evaluation,
It is characterized in that, the chemotherapy refers to docetaxel and prednisone combination therapy.
3. the reagent for being detected to combined mark object described in claim 1.
4. reagent according to claim 3, which is characterized in that refer to for being detected to the expression quantity for combining marker
Reagent.
5. reagent according to claim 3, which is characterized in that include in the reagent：
Primer pair for expanding P53 genes, nucleotide sequence is as shown in SEQ ID NO. 1~2；
Primer pair for expanding GSTP1 genes, nucleotide sequence is as shown in SEQ ID NO. 5~6；
For the primer of miRNA-155 reverse transcriptions, nucleotide sequence is as shown in SEQ ID NO. 34；
Primer pair for expanding miRNA-155 reverse transcriptions cDNA, nucleotide sequence is as shown in SEQ ID NO. 35~36；
Primer pair for expanding reference gene U6, nucleotide sequence is as shown in SEQ ID NO. 37~38；
Primer pair for expanding reference gene GAPDH, nucleotide sequence is as shown in SEQ ID NO. 39~40.
6. including the kit of the reagent described in claim 3.
7. the reagent described in claim 3 is in preparing Hormone refractory prostate cancer chemotherapy resistance recruitment evaluation kit
Using.
8. application according to claim 7, which is characterized in that refer to whether assessing the value of following formula F more than 3.2, work as F
Value be more than 3.2 when, it is believed that there are drug resistance risks：
F1=((- Ln (expression quantity of the miR-155 relative to U6))-0.5+ 0.71) × (Ln (expression quantity of the TP53 relative to GAPDH)+
0.24) -0.52 × Ln (expression quantity of the GSTP1 relative to GAPDH).
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2017102156592 | 2017-04-04 | ||
| CN201710215659.2A CN106834526A (en) | 2017-04-04 | 2017-04-04 | A kind of Polymorphism mark and its detection kit for prostate cancer therapy assessment |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108486249A true CN108486249A (en) | 2018-09-04 |
Family
ID=59142008
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710215659.2A Withdrawn CN106834526A (en) | 2017-04-04 | 2017-04-04 | A kind of Polymorphism mark and its detection kit for prostate cancer therapy assessment |
| CN201810287522.2A Withdrawn CN108486249A (en) | 2017-04-04 | 2018-03-31 | A kind of Polymorphism marker and its detection kit for prostate cancer therapy assessment |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710215659.2A Withdrawn CN106834526A (en) | 2017-04-04 | 2017-04-04 | A kind of Polymorphism mark and its detection kit for prostate cancer therapy assessment |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN106834526A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112195224A (en) * | 2020-10-23 | 2021-01-08 | 漯河医学高等专科学校 | Application of gene combined detection reagent in immunotherapy of lung cancer patients |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2008135923A (en) * | 2008-09-05 | 2010-03-10 | Государственное учреждение Главный клинический военный госпиталь ФСБ России (RU) | METHOD FOR EARLY DIAGNOSIS OF PROSTATE CANCER USING MOLECULAR GENETIC ONCOMARKERS |
| CN103025890A (en) * | 2010-04-06 | 2013-04-03 | 卡里斯生命科学卢森堡控股 | Circulating biomarkers for disease |
| RU2012141174A (en) * | 2012-09-25 | 2014-03-27 | Михаил Иосифович Коган | METHOD FOR DIFFERENTIAL DIAGNOSTICS OF PROSTATE CANCER |
-
2017
- 2017-04-04 CN CN201710215659.2A patent/CN106834526A/en not_active Withdrawn
-
2018
- 2018-03-31 CN CN201810287522.2A patent/CN108486249A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2008135923A (en) * | 2008-09-05 | 2010-03-10 | Государственное учреждение Главный клинический военный госпиталь ФСБ России (RU) | METHOD FOR EARLY DIAGNOSIS OF PROSTATE CANCER USING MOLECULAR GENETIC ONCOMARKERS |
| CN103025890A (en) * | 2010-04-06 | 2013-04-03 | 卡里斯生命科学卢森堡控股 | Circulating biomarkers for disease |
| RU2012141174A (en) * | 2012-09-25 | 2014-03-27 | Михаил Иосифович Коган | METHOD FOR DIFFERENTIAL DIAGNOSTICS OF PROSTATE CANCER |
Non-Patent Citations (1)
| Title |
|---|
| ZHI‑KANG CAI ET AL.: "microRNA‑155 promotes the proliferation of prostate cancer cells by targeting annexin 7", 《 MOLECULAR MEDICINE REPORTS》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112195224A (en) * | 2020-10-23 | 2021-01-08 | 漯河医学高等专科学校 | Application of gene combined detection reagent in immunotherapy of lung cancer patients |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106834526A (en) | 2017-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107849606A (en) | The method for improving sequencing sensitivity of future generation | |
| CN106834470A (en) | Purposes of the miRNA in cancer diagnosing kit is prepared | |
| CN110241208A (en) | Application of the TREM2 as the molecular marker of early diagnosis coronary heart disease | |
| CN107475441B (en) | Biomarker for predicting responsiveness of breast cancer patient to AT regimen neoadjuvant chemotherapy | |
| CN108624693A (en) | Applications of the miR-577 in preparing diagnosis of nephropathy marker | |
| CN109022583A (en) | Hsa_circ_0021977 is preparing the application on Diagnosis of Breast cancer product | |
| CN108486249A (en) | A kind of Polymorphism marker and its detection kit for prostate cancer therapy assessment | |
| CN109536612B (en) | Plasma miRNA marker related to nasopharyngeal carcinoma auxiliary diagnosis and application thereof | |
| CN114015767B (en) | Serum circRNA marker for identifying craniosynostosis and application thereof | |
| CN110656171A (en) | Application of small nucleolus ribonucleic acid SNORD33 as biomarker for preparing detection kit | |
| KR102083956B1 (en) | Biomarkers for prediciting prognosis of lenalidomide plus dexamethasone treatment in patients with multiple myeloma | |
| CN106148562A (en) | For detecting the label of carcinoma of prostate | |
| CN110229891A (en) | The product of non-invasive diagnosis Male Osteoporosis | |
| CN107130028B (en) | Applications of the SYCE3 in diagnosis of colorectal carcinoma and treatment effectiveness evaluation | |
| CN114746551A (en) | Marker for diagnosing colorectal cancer, method for assisting diagnosis of colorectal cancer, method for collecting data to be used for diagnosis of colorectal cancer, diagnostic kit for colorectal cancer, therapeutic agent for colorectal cancer, method for treating colorectal cancer, and method for diagnosing colorectal cancer | |
| CN105950714B (en) | It is a kind of diagnose osteoarthritis product and its application | |
| CN105255865A (en) | Fluorescent quantitative PCR (polymerase chain reaction) detection kit for TYMS (thymidylate synthase) gene expression quantity and application of kit | |
| Björck et al. | Blood cell telomere length among patients with an isolated popliteal artery aneurysm and those with multiple aneurysm disease | |
| CN109371023A (en) | A kind of circular rna hsa_circKIAA1199_006 and its specificity amplification primer and application | |
| CN113186285B (en) | Method for auxiliary diagnosis of gastric cancer and miRNA combination used in method | |
| US11676724B1 (en) | Method of calculating diagnostic score for prostate cancer and use thereof | |
| CN108753956A (en) | Application of the KDSR genes in preparing myocardial infarction diagnosis tool | |
| EP4212632A1 (en) | Method for calculating prostate cancer diagnostic score, and uses thereof | |
| CN107177674A (en) | SPHAR as abdominal aneurvsm diagnosis and treatment target | |
| CN106566886A (en) | Reagent for prognosis assessment of pancreatic cancers, and application of reagent |
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 | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20180904 |