CN105349665B - Kit of colorectal cancer risk of recurrence and application thereof after assessment oxaliplatin chemotherapeutic - Google Patents
Kit of colorectal cancer risk of recurrence and application thereof after assessment oxaliplatin chemotherapeutic Download PDFInfo
- Publication number
- CN105349665B CN105349665B CN201510852099.2A CN201510852099A CN105349665B CN 105349665 B CN105349665 B CN 105349665B CN 201510852099 A CN201510852099 A CN 201510852099A CN 105349665 B CN105349665 B CN 105349665B
- Authority
- CN
- China
- Prior art keywords
- seq
- sequence
- primer
- primer pair
- gene
- 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.)
- Active
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
- C12Q2600/118—Prognosis of disease development
Abstract
Kit the present invention relates to colorectal cancer risk of recurrence after assessment oxaliplatin chemotherapeutic and application thereof.Including expanding the first primer pair of reference gene GAPDH, to the second primer pair of amplification gene PTK2, to the third primer pair of amplification gene WNK1, to the 4th primer pair of amplification gene STK39, to the 5th primer pair of amplification gene DNAJC6, to the 6th primer pair of amplification gene NME4, to the 7th primer pair of amplification gene BRD2, to the 8th primer pair of amplification gene PPP3R1, to the 9th primer pair of amplification gene EYA4, to the tenth primer pair of amplification gene STK32A.The kit of the present invention can be used for evaluating assessment of the colorectal cancer patients using risk of recurrence after the chemotherapeutics of induced DNA damage.
Description
[technical field]
The present invention relates to drug test kits, and in particular to colorectal cancer risk of recurrence after assessment oxaliplatin chemotherapeutic
Kit and application thereof.
[background technology]
Colorectal cancer is one of most commonly seen malignant tumour, and the incidence ascendant trend of CHINESE REGION is fairly obvious, position
Occupy whole malignant tumour third positions, serious threat human health.
With the development of molecular analysis methods, various new diagnosis and treatment methods and new drug continue to bring out, colorectal cancer
Therapeutic effect has clear improvement.However, although traditional clinical pathological factors can provide certain treatment for colorectal cancer patients
Information is imitated, but these risk factors cannot be quantized and integrate well the degree of colorectal cancer influence prognosis, therefore for
Individual patients, current therapy and the medication selection still traditional histopathology of heavy dependence and clinical examination, it is right
The evaluation accuracy of curative effect is not high.In this regard, clinical investigators propose various prognosis and curative effect individuation forecasting tool, to
Reference is provided for colorectal cancer patients postoperative chemotherapy medication by simple and effective Forecasting Methodology, so as to improve the standard of chemotherapy medication
True property improves quality of life of patients.These forecasting tools include gene copy number variation analysis.
At present about gene copy number variation and tumor prognosis correlation research it has been reported that genome 16p13.3,
The copy number variation in the sites such as 1p36 and 4q22.1-4q35.2 is related to the prognosis recurrence of II-III phase colorectal cancers
(Mampaey E,Fieuw A,Van Laethem T,et al.Focus on 16p13.3 Locus in Colon
Cancer.PLoS One.2015 Jul 29；10(7):e0131421；Mayrhofer M,Kultima HG,Birgisson
H,et al.1p36 deletion is a marker for tumour dissemination in microsatellite
stable stage II-III coloncancer.BMC Cancer.2014 Nov 24；14:872；Smith DH,
Christensen IJ,Jensen NF,et al.An explorative analysis of ERCC1-19q13 copy
number aberrations in a chemonaive stage III colorectal cancer cohort.BMC
Cancer.2013 Oct 21；13:489；Brosens RP,Belt EJ,Haan JC,et al.Deletion of
chromosome 4q predicts outcome in stage II colon cancer patients.Cell Oncol
(Dordr).2011 Jun；34(3):215-23).However, suggest as American National synthesis cancer network how-tos (NCCN)
III phase colorectal cancer patients are in the first-line drug oxaliplatin through the postoperative adjuvant chemotherapy of radical excision, and but there is no effectively at present
Forecasting tool be supplied to clinician for distinguish may benefit in oxaliplatin adjuvant chemotherapy method it is more or benefit it is less
PATIENT POPULATION, so as to CRC patient carry out personalized drug therapy.This reality seriously limits oxaliplatin in the III phases
The promotion of curative effect in post operative colo-rectal cancer adjuvant chemotherapy, 35% III phases colorectal cancer patients can not be auxiliary from postoperative oxaliplatin
Help benefit (Uncu D, Aksoy S, Cetin B, et al.Results of adjuvant FOLFOX regimens in chemotherapy
in stage III colorectal cancer patients:retrospective analysis of
667patients.Oncology.2013；84(4):240-245).Therefore, the clinical treatment of colorectal cancer be badly in need of one kind can
The easy method for accurately carrying out prognosis and oxaliplatin outcome prediction, important ginseng is provided for colorectal cancer patients postoperative
It examines.
Protein kinase is sent out in the physiology courses such as the metabolism of the generation of tumour, progress and tumour cell and the generation of drug resistance
Important role is waved, therefore can be as evaluation and test tumor prognosis and the evaluation metrics of chemotherapeutic efficacy.The present invention is in 98 parts of III phases
Post operative colo-rectal cancer oxaliplatin adjuvant chemotherapy is suffered from
The copy number of 626 protein kinase genes is detected in person's tumor tissues, finds wherein 9 protein kinase gene copies
Number variation is closely related with colorectal cancer curative effect.9 genes are divided by following groups according to functional clustering：
| Gene Symbol | Functional Categories |
| BRD2 | Wnt/Hedgehog/Notch signaling |
| PTK2 | Protein tyrosine kinase signaling |
| DNAJC6 | Protein tyrosine phosphatase |
| EYA4 | Protein phosphatase |
| WNK1 | Serine/threonine protein kinase |
| STK32A | Serine/threonine protein kinase |
| STK39 | MAPK signaling |
| PPP3R1 | MAPK signaling |
| NME4 | Metabolic regulation |
Wherein, BRD2 belongs to BET (bromodomainsandextraterminaldomain) protein family, has transcription
Regulatory factor activity, participate in kinds of tumors gene such as c-MYC, BCL2 and SP1 transcription (Fu LL, Tian M, LiX,
etal.InhibitionofBET bromodomains as a therapeutic strategy for cancer drug
discovery.Oncotarget.2015 Mar 20；6(8):5501-16；Minghua Wu,Xiayu Li,Xiaoling
Li,et al.Signaling Transduction Network Mediated by Tumor Suppressor/
Susceptibility Genes in NPC.Curr Genomics.2009 Jun；10(4):216–222).
PTK2 belongs to the FAK subfamily of protein tyrosine kinase family.It participates in multiple signal paths and includes PI3K/
Activation (Sun L, Liu L, the Liu X, et of AKT, MAPK/ERK and GTPase-activating proteins
al.MGr1-Ag/37LRP induces cell adhesion-mediated drug resistance through FAK/
PI3K and MAPK pathway in gastric cancer.Cancer Sci.2014Jun；105(6):651-9；
Hecker TP,Ding Q,Rege TA,et al.Overexpression of FAK promotes Ras activity
through the formation of a FAK/p120RasGAP complex in malignant astrocytoma
cells.Oncogene.2004 May 13；23(22):3962-71), important work is played in the proliferation of tumour and transfer process
With.
DNAJC6 belongs to the DNAJ/HSP40 families of evolution conservative, by activating work of the ATPsae come regulatory molecule companion
Property.GO functional analyses show its activity with Protein-tyrosine-phosphatase.Research in liver cancer finds that DNAJC6 passes through EMT
Regulate and control transfer (Yang T, Li XN, Li XG, the et al.DNAJC6 promotes hepatocellular of liver cancer cells
carcinoma progression through induction of epithelial-mesenchymal
transition.Biochem Biophys Res Commun.2014 Dec 12；455(3-4):298-304.)
EYA4 encodes EYA families (eyesabsentfamily) albumen.EYA4 has protein phosphatase activity, can
It is played a role by the transcription of controlling gene, functional analysis shows that EYA4 participates in DNA and repairs, and apoptosis energy physiology course prompts it
It may participate in the pathologic process of tumour.Research shows that there are Regulation by Methylation in kinds of tumors by EYA4, it is thin in tumour to influence it
Expression (Hou X, PengJX, Hao XY, et al.DNA methylation profiling identifies in born of the same parents
EYA4 gene as a prognostic molecular marker in hepatocellular carcinoma.Ann
Surg Oncol.2014 Nov；21(12):3891-9；Kisiel JB,Garrity-Park MM,Taylor WR,et
al.Methylated eyes absent 4(EYA4)gene promotor in non-neoplastic mucosa of
ulcerative colitis patients with colorectal cancer:evidence for a field
effect.nflamm Bowel Dis.2013 Sep；19(10):2079-83.), the research of Kim SJ et al. finds EYA4
Expression deletion in colon cancer, and prove that its tumor suppressor gene new as one takes part in the pathologic process (Kim of colon cancer
SJ,Tae CH,Hong SN,et al.EYA4Acts as a New Tumor Suppressor Gene in Colorectal
Cancer.Mol Carcinog.2015 Dec；54(12):1748-57.).
WNK1 belongs to a subfamily (WNKfamily) of Serine/threonine protease families.WNK families include
4 members, WNK1-4.More and more researchs confirm that WNK protein kinases family takes part in the grade of many A signal pathways in tumour
Join amplification process, Moniz S, Jordan P's research shows that WNK in tumour cell cycle, metastases and apoptosis of tumor cells
In play a significant role (Moniz S, Jordan P.Emerging roles for WNK kinases in cancer.Cell
Mol Life Sci.2010 Apr；67(8):1265-76.).
The albumen of STK32A gene codes belongs to Serine/threonine protein kinases family, participates in intracellular WNT3A
The regulation and control of post-stimulatory Wnt/beta-catenin signal paths.
One Serine/threonine kinases of STK39 gene codes, by MAPK signal paths participate in cell stress
Reaction.The research of Qi H et al. thinks expression of the STK39 in prostate gland cancer cell there are Androgen-dependent, and androgen lures
The STK39/MAPK signal cascades led react may play a significant role in prostate cancer (QiH, Labrie Y, Grenier
J,et al.Androgens induceexpression of SPAK,a STE20/SPS1-related kinase,in
LNCaP human prostatecancer cells.Mol Cell Endocrinol.2001 Sep；182(2):181-
92.).Performance key effect in the Hela Cell Apoptosis that the shearing of STK39 and inactivation induces in TRAIL (Polek TC,
Talpaz M,Spivak-Kroizman TR.TRAIL-induced cleavage and inactivation of SPAK
sensitizes cells to apoptosis.Biochem Biophys Res Commun.2006 Oct 27；349(3):
1016-24.)。
Adjusting subunit of the albumen of PPP3R1 gene codes as calcineurin is that a calcium ion relies on, can be by calcium
The phosphoprotein phosphatase of adjusting phosphatase activation, functional analysis system show that PPP3R1 is related to PI3K and MAPK signal paths.
NME4 belongs to nucloside-diphosphate kinase family, and main function is synthesis nucleoside triphosphate, participates in the metabolism of cell
Journey.Multiple studies have shown that there is the phenomenon that being overexpressed in NME4 in different tumor types.Seifert Metal pass through RT-
The expression that PCR and immunohistochemistry have detected NME families in colon cancer and gastric cancer finds the expression of NME4 and the TNM stage phase of tumour
It closes, research thinks that NME4 participates in development (Seifert M, Welter C, the Mehraein Y, et of gastric cancer and colon cancer
al.Expression of the nm23 homologues nm23-H4,nm23-H6,and nm23-H7 in human
gastric and coloncancer.J Pathol.2005 Apr；205(5):623-32.).
Therefore, the copy number of the invention by detecting above-mentioned 9 genes calculates summary and show that a set of assessment oxaliplatin is treated
The risk of recurrence methods of marking of effect effectively can provide important references for colorectal cancer patients postoperative.Further, originally
Invention, using designed, designed and the internal reference optimized and purpose primer, incorporates real time fluorescent quantitative using fluorescent quantitative PCR technique
Detection kit is made in PCR reagent, and clinical detection is facilitated to use.
[invention content]
The primary and foremost purpose of the present invention is found with recurring relevant important gene after colon cancer chemotherapy, next is to provide one
For the method for recurring risk assessment after colorectal carcinoma chemotherapy, then made by one group of important gene after detecting colon cancer chemotherapy
The kit of recurring risk assessment easy can rapidly evaluate patient if appropriate for progress chemotherapy.
To achieve these goals, the present invention provides a kind of examination of colorectal cancer risk of recurrence after assessment oxaliplatin chemotherapeutic
Agent box, including:
A. it is SEQ ID NO by sequence：1 sense primer and sequence is SEQ ID NO：The of 2 downstream primer composition
One primer pair；
B. it is SEQ ID NO by sequence：3 sense primer and sequence is SEQ ID NO：The of 4 downstream primer composition
Two primer pairs；
C. it is SEQ ID NO by sequence：5 sense primer and sequence is SEQ ID NO：The of 6 downstream primer composition
Three-primer pair；
D. it is SEQ ID NO by sequence：7 sense primer and sequence is SEQ ID NO：The of 8 downstream primer composition
Four primer pairs；
E. it is SEQ ID NO by sequence：9 sense primer and sequence is SEQ ID NO：The of 10 downstream primer composition
Five primer pairs；
F. it is SEQ ID NO by sequence：11 sense primer and sequence is SEQ ID NO：12 downstream primer composition
6th primer pair；
G. it is SEQ ID NO by sequence：13 sense primer and sequence is SEQ ID NO：14 downstream primer composition
7th primer pair；
H. it is SEQ ID NO by sequence：15 sense primer and sequence is SEQ ID NO：16 downstream primer composition
8th primer pair；
I. it is SEQ ID NO by sequence：17 sense primer and sequence is SEQ ID NO：18 downstream primer composition
9th primer pair；
J. it is SEQ ID NO by sequence：19 sense primer and sequence is SEQ ID NO：20 downstream primer composition
Tenth primer pair.
Mentioned reagent box can be used for the reagent of risk of recurrence after preparation assessment colorectal carcinoma chemotherapy.
Because the active material that main anti-tumor is played in FOLFOX chemotherapy regimens is 5-FU (5FU) and oxaliplatin,
Their mode of action is induced DNA damage so as to cause Apoptosis, and therefore, the present invention relates to one group of important genes as commenting
Valency colorectal cancer patients using induced DNA damage chemotherapeutics after recurring risk assessment index.
Because DNA copy number reduction directly results in gene expression reduction, therefore, the present invention also relates to utilize one group of important base
Occur and after colorectal cancer patients take chemotherapeutics as evaluation colorectal cancer because expressing (including mRNA and protein level)
The index of recurrence.
[description of the drawings]
Fig. 1 is the Relative copy number of 9 genes and the relation schematic diagram of risk of recurrence after colorectal cancer patients chemotherapy；
Fig. 2 evaluates colorectal cancer patients chemotherapy recurring risk assessment schematic diagram for 9 genes.
[specific embodiment]
The Relative copy number that the present invention detects 9 genes in colorectal cancer sample by fluorescent quantitative PCR technique changes, and
Whether there is significant difference with the copy number variation of 9 genes of student t test and judges；Then, colorectal cancer patients are carried out
Life cycle count, by patient be divided into less than 5 years with 5 years or more two groups, with two groups of the student t test and judges gene of crowd 9
The whether significant difference of Relative copy number.Patient is analyzed by the multifactor homing method of cox models with SPSS softwares later to answer
The relationship of the Relative copy number of hair and 9 genes, establishes the calculation formula of recurrence scoring (RS), and survival rate is assessed using K-M methods,
The difference of survival rate between different groups is examined using log-rank.And with cox regression analyses specificity and sensitivity.
It is specific embodiments of the present invention below：
The present embodiment includes the detection primer of 10 couples of qPCR
A. it is SEQ ID NO by sequence：1 sense primer and sequence is SEQ ID NO：The of 2 downstream primer composition
One primer pair, to expand reference gene GAPDH.
B. it is SEQ ID NO by sequence：3 sense primer and sequence is SEQ ID NO：The of 4 downstream primer composition
Two primer pairs, to amplification gene PTK2.
C. it is SEQ ID NO by sequence：5 sense primer and sequence is SEQ ID NO：The of 6 downstream primer composition
Three-primer pair, to amplification gene WNK1.
D. it is SEQ ID NO by sequence：7 sense primer and sequence is SEQ ID NO：The of 8 downstream primer composition
Four primer pairs, to amplification gene STK39.
E. it is SEQ ID NO by sequence：9 sense primer and sequence is SEQ ID NO：The of 10 downstream primer composition
Five primer pairs, to amplification gene DNAJC6.
F. it is SEQ ID NO by sequence：11 sense primer and sequence is SEQ ID NO：12 downstream primer composition
6th primer pair, to amplification gene NME4.
G. it is SEQ ID NO by sequence：13 sense primer and sequence is SEQ ID NO：14 downstream primer composition
7th primer pair, to amplification gene BRD2.
H. it is SEQ ID NO by sequence：15 sense primer and sequence is SEQ ID NO：16 downstream primer composition
8th primer pair, to amplification gene PPP3R1.
I. it is SEQ ID NO by sequence：17 sense primer and sequence is SEQ ID NO：18 downstream primer composition
9th primer pair, to amplification gene EYA4.
J. it is SEQ ID NO by sequence：19 sense primer and sequence is SEQ ID NO：20 downstream primer composition
Tenth primer pair, to amplification gene STK32A.
1st, experimental subjects：
The research object of the present embodiment selects the colorectal cancer collected during 2006 in December, 2011 in Ruijin Hospital to suffer from
Person's sample by data to arrange and follow-up, therefrom has chosen 98 colorectal cancer tumour paraffin-embedded tissue samples.
It is included in and exclusion criteria：
(1) the equal initial diagnosis of all cases received for colorectal cancer and not radiation and chemotherapy before.
(2) in order to reduce inter-sample difference to the greatest extent, we only study the III phase large intestines performed the operation through radical excision
Cancer patient, and at least six period is used using oxaliplatin as a line adjuvant chemotherapy drug, all patients are according to the U.S.
Cancer Joint Committee tumour node transfer Staging System (TNM) carries out by stages.
(3) without other organ tumor medical histories；It is difficult lower family history：Without colorectal carcinoma history, no adenoma in 1~2 grade of relatives
Property polyp medical history and Familial Occurrence syndrome history, mainly including familial adenomatous polyposis (FAP), hereditary nonpolyposis
Colorectal cancer (HNPCC) etc.；
(4) we have looked back the medical records of patient and have collected clinical diagnosis information, including diagnosis date, clinical stage,
Knub position, histology by stages, pathological staging and treatment information.Patient's recurrence, dead information by medical records and phone with
It visits and obtains, follow-up in January, 2014.
2nd, experimental method：
(1) using the tumor tissues sample of above-mentioned 98 colorectal cancer patients underwent operatives excision, the sample of each paraffin embedding
Originally 50-100 slices are cut, are passed through after haematoxylin ＆ eosin (HE) dyeing by 3 pathologists diagosis.Three pathology doctors are scraped by hand
The tumour cell area that teacher unanimously thinks is more than the slice of slice area 70%.Tumor tissue section, which is immersed in dimethylbenzene, to dewax,
With the QIAamp-DNA-FFPE-Tissue Kit of Qiagen companies, according to the explanation of kit, the DNA samples in tissue are extracted
This.DNA sample after extracting is stored in -20 DEG C of low temperature refrigerator.
(2) with the concentration and matter of the DNA of Thermo Scientific Varioskan Flash microplate reader detection extracting
Amount.
(3) 60 genes and internal reference in 98 parts of DNA samples are detected with the iQ5 quantitative PCR apparatus that Bio-rad companies produce
The copy number of GAPDH genes.Each sample draws 20ng genomic DNAs, using SYBR Green kits (Qiagen,
German quantitative PCR) is carried out, the primer is shown in Table 1.PCR programs are：95 DEG C of denaturation 15s, 58 DEG C of primer annealing 20s, 72 DEG C are prolonged
Stretch 20s.
3rd, interpretation of result
(1) each sample sets three multiple holes, if different more than one cycle of multiple holes Ct value differences, will be removed out experiment point
Analysis.Following (Livak K, the Schmittgen TD.Analysis of relative gene of copy number analytic process
expression data using real-time quantitative CR and the 2(-DeltaDelta C(T))
method.Methods 2001；25:402–8.)：1st, the average value that each sample target gene three repeats Ct values is calculated, simultaneously
Calculate the average value that corresponding reference gene three repeats；2nd, the average value of all sample target gene and reference gene is calculated；3rd, it uses
The target gene Average Ct values of each sample subtract the Average Ct values of all sample target gene, and as target gene △ Ct are interior
Join the similary algorithm process of △ Ct；4th, the 2 of each gene is calculated-ΔCt(target gene) also calculates the 2 of reference gene-ΔCt(internal reference base
Cause)；Target gene 2-ΔΔCt=2Δ Ct (target gene)/- Δ Ct (reference gene)。
(2) we are using 60 contacting between gene copy number and patient survival of multifactor Cox model evaluations, sieve
Selecting 9 genes and life cycle after colorectal cancer patients chemotherapy has significant correlation.(being shown in Table 2) is according to each copy number changes
After detected value carries out weight, the calculation formula of prediction chemotherapy risk of recurrence is obtained.Specific formula for calculation is as follows：RS (recurrence scoring)
=-0.985*PTK2-1.443*WNK1+1.498*STK39-0.388*NME4-0.267*DNAJC 6-1.568*PPP3R1-
0.441*EYA4+0.662*STK32A+0.805*BRD2
Table 2
Multifactor Cox risks regression model
(3) in order to further evaluate the risk of recurrence, and so that kit is easy to use, we are fixed by RS (recurrence is scored)
Justice is：The first kind (recurring low-risk in RS≤5.5511), the second class (5.5511<RS<6.87 recurrence medium risks), third class
(recurring high risk in RS≤6.87) single factor test cox risk regression analyses show that this three classes RS has evaluated 9 gene copies well
Contacting between number variation and risk of recurrence.
(being shown in Table 3)
Table 3
RS classification single factor test cox risk regression models
(4) it assesses gene copy number using Kaplan-Meier survivorship curves and the relationship of life cycle is shown in Fig. 1.Low-risk
Group has preferable prognosis than medium risk group, and medium risk group then has preferable prognosis than high risk group；Patient is through Ao Shali
When reaching identical life span after platinum chemotherapy, the survival probability of low-risk group is apparently higher than medium risk group, and medium risk group
Survival probability be then apparently higher than high risk group, have significant difference between three groupings in model.
(5) RS (recurrence is scored) and life cycle are carried out Receiver operating curve (ROC) to analyze, as shown in Fig. 2, with
In the performance parameter of the RS (recurrence scoring) of assessment colorectal cancer patients recurring risk assessment after oxaliplatin chemotherapeutic be tested
Area (AUC) value below person's performance curve is 0.74, P=0.007, and 95% confidence interval is 0.601-0.879, is more than
0.5, illustrate that RS (recurrence scoring) has very well for assessing risk of recurrence of the colorectal cancer patients after oxaliplatin chemotherapeutic
Application value.
4th, the application in risk of recurrence judgement
The operation excision cancerous tissue sample of colorectal cancer patients to be checked is acquired, according to preceding method, extracts DNA sample, and
Copy number of 9 genes in the specimens DNA sample is detected, then, is calculated according to the calculation formula of risk of recurrence
Go out RS (recurrence scoring), evaluate the patient to be checked if appropriate for application oxaliplatin as chemotherapeutics, provided for clinical application
With reference to.
5th, detection kit
Kit of the present invention is formed by following reagent and (is shown in Table 4), and source is as follows, and kit of the present invention is detected for 100 person-portions
Using -20 DEG C of preservations：
4 kit forms of table
| Ingredient | Volume |
| 2*SYBR Green | 10μl |
| Primer Mix | 20ng(2.5μl) |
| Template | 20ng(2.5μl) |
| Ultra-pure water | 5μl |
| Total volume | 20ul |
Claims (2)
1. a kind of kit of colorectal cancer risk of recurrence after assessment oxaliplatin chemotherapeutic, it is characterised in that including:
A. it is SEQ ID NO by sequence：1 sense primer and sequence is SEQ ID NO：The first of 2 downstream primer composition is drawn
Object pair；
B. it is SEQ ID NO by sequence：3 sense primer and sequence is SEQ ID NO：The second of 4 downstream primer composition is drawn
Object pair；
C. it is SEQ ID NO by sequence：5 sense primer and sequence is SEQ ID NO：The third of 6 downstream primer composition is drawn
Object pair；
D. it is SEQ ID NO by sequence：7 sense primer and sequence is SEQ ID NO：The 4th of 8 downstream primer composition draws
Object pair；
E. it is SEQ ID NO by sequence：9 sense primer and sequence is SEQ ID NO：The 5th of 10 downstream primer composition draws
Object pair；
F. it is SEQ ID NO by sequence：11 sense primer and sequence is SEQ ID NO：The 6th of 12 downstream primer composition
Primer pair；
G. it is SEQ ID NO by sequence：13 sense primer and sequence is SEQ ID NO：The 7th of 14 downstream primer composition
Primer pair；
H. it is SEQ ID NO by sequence：15 sense primer and sequence is SEQ ID NO：The 8th of 16 downstream primer composition
Primer pair；
I. it is SEQ ID NO by sequence：17 sense primer and sequence is SEQ ID NO：The 9th of 18 downstream primer composition
Primer pair；
J. it is SEQ ID NO by sequence：19 sense primer and sequence is SEQ ID NO：The tenth of 20 downstream primer composition
Primer pair.
2. a kind of purposes of kit described in claim 1 after assessment colorectal carcinoma chemotherapy is prepared in risk of recurrence reagent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510852099.2A CN105349665B (en) | 2015-11-27 | 2015-11-27 | Kit of colorectal cancer risk of recurrence and application thereof after assessment oxaliplatin chemotherapeutic |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510852099.2A CN105349665B (en) | 2015-11-27 | 2015-11-27 | Kit of colorectal cancer risk of recurrence and application thereof after assessment oxaliplatin chemotherapeutic |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105349665A CN105349665A (en) | 2016-02-24 |
| CN105349665B true CN105349665B (en) | 2018-06-19 |
Family
ID=55325725
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510852099.2A Active CN105349665B (en) | 2015-11-27 | 2015-11-27 | Kit of colorectal cancer risk of recurrence and application thereof after assessment oxaliplatin chemotherapeutic |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105349665B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112680517B (en) * | 2020-12-28 | 2023-06-02 | 广东南芯医疗科技有限公司 | Guiding method and kit for oxaliplatin personalized medicine genes |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004094636A1 (en) * | 2003-04-24 | 2004-11-04 | Galapagos Genomics N.V. | Effective sirna knock-down constructs |
| WO2009071720A1 (en) * | 2007-12-04 | 2009-06-11 | Universidad Autónoma de Madrid | Genomic imprinting for the prognosis of the course of colorectal adenocarcinoma |
| WO2011091272A1 (en) * | 2010-01-21 | 2011-07-28 | Dana-Farber Cancer Institute, Inc. | Context specific genetic screen platform to aid in gene discovery and target validation |
| CN103710451A (en) * | 2013-12-26 | 2014-04-09 | 上海锐赛生物技术有限公司 | Application of PIK3C2G in evaluation and detection kit for curative effect of colorectal cancer chemotherapy |
-
2015
- 2015-11-27 CN CN201510852099.2A patent/CN105349665B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004094636A1 (en) * | 2003-04-24 | 2004-11-04 | Galapagos Genomics N.V. | Effective sirna knock-down constructs |
| WO2009071720A1 (en) * | 2007-12-04 | 2009-06-11 | Universidad Autónoma de Madrid | Genomic imprinting for the prognosis of the course of colorectal adenocarcinoma |
| EP2236626A1 (en) * | 2007-12-04 | 2010-10-06 | Universidad Autónoma De Madrid | Genomic imprinting for the prognosis of the course of colorectal adenocarcinoma |
| WO2011091272A1 (en) * | 2010-01-21 | 2011-07-28 | Dana-Farber Cancer Institute, Inc. | Context specific genetic screen platform to aid in gene discovery and target validation |
| CN103710451A (en) * | 2013-12-26 | 2014-04-09 | 上海锐赛生物技术有限公司 | Application of PIK3C2G in evaluation and detection kit for curative effect of colorectal cancer chemotherapy |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105349665A (en) | 2016-02-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Giulietti et al. | Identification of candidate miRNA biomarkers for pancreatic ductal adenocarcinoma by weighted gene co-expression network analysis | |
| Liu et al. | A novel strategy of integrated microarray analysis identifies CENPA, CDK1 and CDC20 as a cluster of diagnostic biomarkers in lung adenocarcinoma | |
| Zhou et al. | Development and validation of an autophagy score signature for the prediction of post-operative survival in colorectal cancer | |
| Glavac et al. | MicroRNAs as novel biomarkers in colorectal cancer | |
| Liu et al. | Identification and validation of gene module associated with lung cancer through coexpression network analysis | |
| Teng et al. | miRNA-200a/c as potential biomarker in epithelial ovarian cancer (EOC): evidence based on miRNA meta-signature and clinical investigations | |
| Singhal et al. | Gene expression profiling of non-small cell lung cancer | |
| Huang et al. | A novel RNA binding protein-related prognostic signature for hepatocellular carcinoma | |
| Chen et al. | Identification of a four hypoxia-associated long non-coding RNA signature and establishment of a nomogram predicting prognosis of clear cell renal cell carcinoma | |
| Lee et al. | Differential expression of microRNAs and their target genes in non-small-cell lung cancer | |
| Zhang et al. | Profile of differentially expressed miRNAs in high-grade serous carcinoma and clear cell ovarian carcinoma, and the expression of miR-510 in ovarian carcinoma | |
| Jiang et al. | Key regulators in prostate cancer identified by co-expression module analysis | |
| Sanchez-Diaz et al. | In silico functional analyses and discovery of survival-associated microRNA signatures in pediatric osteosarcoma | |
| Zhou et al. | Development and validation of a prognostic signature for malignant pleural mesothelioma | |
| Zhang et al. | Recurrence-associated multi-RNA signature to predict disease-free survival for ovarian cancer patients | |
| Kessous et al. | Distinct homologous recombination gene expression profiles after neoadjuvant chemotherapy associated with clinical outcome in patients with ovarian cancer | |
| He et al. | Seven-microRNA panel for lung adenocarcinoma early diagnosis in patients presenting with ground-glass nodules | |
| Cheng et al. | A signature of nine lncRNA methylated genes predicts survival in patients with glioma | |
| Shi et al. | Gene expression signature for detection of gastric cancer in peripheral blood | |
| EP3444362B1 (en) | Circulating mirnas as early detection marker and prognostic marker | |
| Peng et al. | Identification of a novel prognostic signature of genome instability-related LncRNAs in early stage lung adenocarcinoma | |
| Wang et al. | Integrative modeling of multiomics data for predicting tumor mutation burden in patients with lung cancer | |
| Sanz-Pamplona et al. | A taxonomy of organ-specific breast cancer metastases based on a protein–protein interaction network | |
| Dang et al. | Microarray and next-generation sequencing to analyse gastric cancer | |
| CN105349665B (en) | Kit of colorectal cancer risk of recurrence and application thereof after assessment oxaliplatin chemotherapeutic |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20180829 Address after: Room 1501-312, Putuo Chamber of Commerce Building, 66 Shatian Street, Donggang Street, Putuo District, Zhoushan City, Zhejiang Province Patentee after: Zhejiang free trade zone ruicai biological medicine technology Co., Ltd. Address before: 200120 room 225, No. 199 Guo Shou Jing Road, Pudong New Area free trade pilot area, Shanghai Patentee before: Shanghai Rui Sai Bioisystech Co., Ltd |
|
| TR01 | Transfer of patent right |