CN103324846A - Screening method of colorectal cancer treatment prognosis biomarkers - Google Patents

Abstract

The invention discloses a screening method of colorectal cancer treatment prognosis biomarkers. The screening method includes the following steps of: (1) making a primary search strategy for screening the biomarkers; (2) making a literature adopting and exclusion criterion; (3) analyzing data and primarily screening the biomarkers; (4) making evident grades; (5) making a high quality evident evaluation criterion; (6) performing grading retrieval and quality evaluation on biomarker clinic evident; (7) performing data analyzing and statistics on the biomarker clinic evident; (8) screening the prognosis biomarkers. The invention further provides the colorectal cancer treatment prognosis biomarkers obtained through screening according to the screening method, and the prognosis biomarkers include KRAS, TYMS, TYMS, EGFR, UGT1A1*28, DPYD, PIK3CA, ERCC1, PTEN, VEGFR, BRAF, GST P1, XRCC1, MTHFR, MSI and the like.

Description

The screening technique of colorectal cancer disease treatment prognosis biomarker

Technical field

The invention belongs to biological technical field, relate to a kind of screening technique for the treatment of the prognosis biomarker based on the colorectal cancer disease of clinical evidence evaluation criterion.

Background technology

Colorectal cancer is common malignant neoplasm in digestive tract, and the incidence of disease accounts for the 3rd of world's malignant tumour, and its case fatality rate accounts for the 4th.Along with the raising of people's living standard, the M ﹠ M of colorectal cancer still is the trend that rises year by year.It is reported that it is about 4% that China colorectal cancer morbidity number increases year by year, far surpasses 2% world level.Although obtained certain progress to the Study of Etiology of colorectal cancer with based on the complex treatment of excision, radiation therapy and chemotherapy, effect is unsatisfactory, and 5 years survival rates of patient with operation are 50%-60% over nearly 40 years.Along with continuing to bring out for the treatment of colorectal cancer medicine, select which medicine to carry out first-line treatment and second line treatment and depend primarily on the toxic and side effect that single medicine and combined chemotherapy produce.Therefore, whether the patient of clear and definite particular medication has the danger that serious toxicity takes place, and whether the prediction patient can benefit just to seem most important from treatment.Prediction index can make colorectal cancer treatment individuation to a certain extent, so may be a direction of individualized treatment according to the feature selecting therapeutic scheme of colorectal cancer patients individual tumors molecular phenotype.

At present, the relevant biomarker of existing a part of oncotherapy is applied in clinical, U.S. food in 2005 and Drug Administration (FDA) require to add caution at the Irinotecan medicine label, suggestion patient reply UGT1A1 before using Irinotecan carries out genetic test, for UGT1A1 gene unconventionality person, processing that should the medicine decrement.The U.S. state-run comprehensive cancer network (National Comprehensive Cancer Network, NCCN) spelling out the KRAS gene in the clinical guidelines of version in 2009 is oncogene common in the tumor disease, the sudden change of KRAS gene can cause colorectal cancer patients antagonism EGFR class target therapeutic agent to produce resistance, therefore the NCCN guide recommends the colorectal cancer tumor patient before accepting the treatment of EGFR targeted drug, should carry out the KRAS detection in Gene Mutation, determine whether use the EGFR targeted drug as the clinical treatment measure according to the result.Newly-increased in the guide of NCCN2011 version, if consider the fluorouracil single therapy, recommend row mispairing reparation (MMR) to detect.Therefore, the relevant biomarker of screening colorectal cancer personalized medicine, make up the diagnostic criteria of colorectal cancer molecule parting, being used for instructing clinical is the important channel of realizing individualized treatment, it is the specific aim that can improve clinical therapy of tumor, reduce the patient treatment expense, save the precondition of valuable treatment time.

Begin the seventies in 20th century, the clinical research personnel begin to carry out the clinical research of all kinds of relevant biomarkers things of colorectal cancer, yet to same problem because be put to the test object, sample size, the restriction of factors such as area, experimental design, the research that carry out in each laboratory may draw the conclusion of inconsistent or even contradiction.The only wheel commentaries on classics between the researcher of these results of study, very little to the clinical practice influence, invalid measure is still continued to be extensive use of.Simultaneously, annual about the thousands of continual renovation of the clinical research of oncotherapy in types of databases, for example deliver in the period of the 1970-2013 among the MEDLINE that research has 200,000 pieces about colorectal cancer, wherein relevant with biomarker above 50,000 pieces.The clinician obtains informational needs cost great amount of manpower and time from these databases, research and application process that influence greatly finishes rectum cancer treatment prognosis biomarker.This dependence medical practice person individual retrieves, collects and assess existing evidence and applies it to practice and decision-making, is not the effective ways of realizing that fundamental research transforms to clinical research.

Therefore, how in the colorectal cancer clinical research data of magnanimity, to filter out the treatment prognosis biomarker with using value and become a urgent problem.In order to eliminate gap and the obstacle between clinical and fundamental research, advance the development of individualized treatment effectively, the two-way conversion between the clinical and fundamental research of Study on Acceleration.A kind of screening colorectal cancer treatment prognostic markers object space method based on clinical evidence of this research invention, the clinical medicine evidence is collected and put in order to this method successively with evidence quality height order, utilize including in evaluation criterion of clinic study that evidence is analyzed, put in order and processes, finally provide objective conclusion, avoided the research deviation of single research, reduce the screening cost simultaneously, shortened screening time.

Summary of the invention

The technical problem to be solved in the present invention provides a kind of method of screening straight treatment of colon cancer prognosis biomarker, this method is based on clinical evidence, by formulating including in and evaluation criterion of clinical evidence, evidence is screened analysis, arrangement and processing, and final screening obtains colorectal cancer disease treatment prognosis biomarker.

In order to solve the problems of the technologies described above, the invention provides a kind of method of screening straight treatment of colon cancer prognosis biomarker, concrete steps comprise:

1. formulate the search strategy of preliminary screening biomarker:

According to PICOS principle deterministic retrieval word, and the descriptor index of employing cloth woods connector formula, wherein descriptor for some vocabulary in the content in full be equal to lexical or textual analysis word and medical science subject at the word of interior expression medical domain, research type and the publication language of retrieval are unrestricted.Utilize term to comprise MEDLINE at database, EMBASE, Cochrane, TRIP, Wei Pu (CPVIP), Chinese biological medical journals bibliographic data base (CMDisc), the comprehensive searching document in database and the total storehouse of Chinese knowledge resource (CNKI) incomparably.The list of references of document is included in inspection in, if used identical research object, chooses the research of all the identical research objects sample size maximum or that comprise.

2. formulating document adopts and exclusion standard:

Adopt and exclusion standard according to document, screen standard compliant literature summary.Adopt standard: 1. research must be the clinical research of carrying out with artificial research object; 2. research must have the treatment intervention means; 3. research contents must have been explored one or more relevant biomarkers for the treatment of; 4. clinical efficacy must be objective life cycle, drug response rate or bad reaction; 5. only include perspective and retrospective study in.Exclusion standard: the basic medical research that does not 1. have molecule, cell and the animal of direct relation with medical practice; 2. study sample size＜20 examples.

3. the preliminary screening of data analysis and biomarker:

According to standard compliant literature summary, initial analysis literature research type and the patient characteristics, therapeutic scheme, the final result index of measurement effect, the observing time of final result of including research in, analyze the biomarker regularity of distribution of different document quality, different treatment means, different treatment results, different research types, make up the biomarker storehouse.The screening biomarker meets one of following standard in the storehouse: 1. the systematic review document is arranged in the evidence; 2. randomized controlled trial research document is arranged in the evidence; 3. cohort study's quantity of document 〉=5 piece in the evidence.

4. formulate the rank of evidence:

According to the evidence classification of colorectal cancer treatment prognosis biomarker, with evidence classification such as table 1:

Table 1 evidence rank

5. formulate high-quality evidence judgment criteria:

4 grades of different literature research types are formulated the judgment criteria of high-quality evidence with reference to Newcastle-Ottawa Scale standard.

(1) systematic review of randomized controlled trial satisfies: 1. the systematic review research and design has clear and definite search strategy, and searching database comprises MEDLINE and EMBASE; The clear and definite standard of including in is arranged; 2. the original of including in satisfies: diagnosing tumor is clear and definite by stages, and sample size 〉=50 examples is lost visit rate＜20%, the existence effect follow up a case by regular visits to time＞2 year, the research and design method is clear and definite, research mode adopts single blind or double blinding; 3. statistical method is reasonable, and carries out heterogeneity and detect; 4. prognostic indicators such as amalgamation result PFS, recurrence, toxicity are clear and definite, do not have and to deliver bias (p〉0.05).

(2) randomized controlled trial satisfies: 1. diagnosing tumor is clear and definite by stages, sample size 〉=200 examples; 2. lose visit rate＜20%, the existence effect follow up a case by regular visits to time＞2 year; 3. research mode adopts single blind or double blinding; 4. the biomarker object detecting method is clear and definite; 5. prognosis final result indexs such as PFS, recurrence, toxicity are clear and definite; 6. positive findings threshold value definition clear-cut.

(3) systematic review of cohort study's test satisfies: 1. the systematic review research and design has clear and definite search strategy, and searching database comprises MEDLINE and EMBASE; Clear and definite including in and exclusion standard arranged; 2. the original of including in satisfies: sample wide material sources, clear and definite by stages, sample size 〉=300 examples is lost visit rate＜20%, the existence effect follow up a case by regular visits to time＞1 year; 3. statistical method is reasonable, and carries out heterogeneity and detect; 4. prognostic indicators such as amalgamation result PFS, recurrence, toxicity are clear and definite, do not have and to deliver bias (p〉0.05).

(4) cohort study test is satisfied: 1. sample wide material sources, sample size 〉=300 examples; 2. diagnosing tumor is clear and definite by stages; 3. lose visit rate＜20%, the existence effect follow up a case by regular visits to time＞2 year; 4. the research and design method is reasonable, and the biomarker object detecting method is clear and definite; 5. prognosis final result indexs such as PFS, recurrence, toxicity are clear and definite; 6. positive findings threshold value definition clear-cut; 7. deliver periodical IF＞2.

6. the grading search of biomarker clinical evidence and quality evaluation:

Evidence rank 1a＞1b＞2a＞2b is comply with in the retrieval of biomarker clinical evidence, with the biomarker relevant clinical evidence in the sequential search primary dcreening operation of the systematic review of randomized controlled trial, randomized controlled trial, systematic review, all correlative study retrieval types, include in and meet document and adopt clinical evidence with exclusion standard, according to high-quality evidence judgment criteria the clinical evidence of including in is carried out quality evaluation again.Evidence quality evaluation process is independently finished by two valuation officers, decides through discussion or transfer to third party's decision if meet the both sides that have different views.

7. the data analysis of biomarker clinical evidence and statistics:

Only the high-quality clinical evidence of biomarker is carried out data analysis and statistics, as evidence 1a, when having only one piece of systematic review in the 2a class, this evidence directly is included into.When many pieces of systematic reviews occurring: (1) includes all systematic reviews in if the result is significantly consistent or unanimity is not remarkable; When (2) result is inconsistent, need to consider that single piece of systematic review includes differences such as the scheme of the original scope of research and quantity, test design, study population's feature, statistical method in, the systematic review of selecting to include in satisfies standard: 1. sample is more; 2. test design more reasonable (including quality analysis, subgroup analysis, sensitivity analysis etc. if any research in).

Evidence 1b, when having only one piece of systematic review among the 2b, this evidence directly is included into.When many pieces of researchs occurring, use meta to analyze amalgamation result, Q-test estimates heterogeneous, and searches heterogeneous source with the subgroup analysis, delivers bias with Begg ' s funnel figure check.The amalgamation result index is included the evidence storehouse in.

8. treatment prognostic markers thing screening:

Prognostic indicators such as The selection result OS, PFS, recurrence, toxicity significantly (p＜0.05) from the evidence of including in do not have the biomarker of delivering bias (p〉0.05).

The present invention also provide simultaneously utilize said method screening and colorectal cancer disease treatment prognosis biomarker, for following any one:

KRAS、TYMS、TYMS、EGFR、UGT1A1*28、DPYD、PIK3CA、ERCC1、PTEN、VEGFR、BRAF、GST?P1、XRCC1、MTHFR、MSI、ERCC2。

Technical advantage of the present invention is:

1. this method has been started the research document of retrieving the mark of correlation thing according to the sequencing of evidence quality grade height successively, can maximally utilise existing research evidence in the data of magnanimity, has shortened screening time, reduces the screening cost.

2. the present invention relates to including in and evaluation criterion of a cover clinic study, utilize this standard that evidence is analyzed, put in order and processes, finishing screen is selected colorectal cancer treatment prognosis biomarker.This standard can have been avoided the final result deviation of single research, can synthetically assess simultaneously the value of biomarker again, makes things convenient for researchist, doctor, enterprise to the practical application of selection markers thing.

Description of drawings

Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.

Fig. 1 is biomarker clinical evidence data screening process flow diagram.

Embodiment

In order to be illustrated more clearly in the present invention, specifically describe the screening technique that colorectal cancer is treated the prognosis biomarker according to Fig. 1 and case study on implementation below.

The present invention is example with ERCC1 gene C 118T polymorphism as the screening process of oxaliplatin treatment colorectal cancer prognosis biomarker, introduces concrete mode of the invention process.

The screening technique of embodiment 1, rectum treatment of cancer prognosis biomarker, carry out following steps successively:

1. formulate the search strategy of primary dcreening operation biomarker:

Sieve and look for colorectal cancer treatment relevant biomarkers thing, according to PICOS principle deterministic retrieval word, the term content comprises: P: straight colon cancer patient; I: medicine; C: no medicine; O: survival value, bad reaction; S: do not limit; Biomarker.The database that retrieval is used comprises MEDLINE, EMBASE, Cochrane, TRIP, Wei Pu (CPVIP), Chinese biological medical journals bibliographic data base (CMDisc), the comprehensive searching document in database and the total storehouse of Chinese knowledge resource (CNKI) incomparably.Be 1980-2012.12.30 retrieval time.

foreign language database retrieval formula: (" rectal ", OR, " rectum ", OR, " colon ", OR, " colonic ", OR, " colorectal ", OR, " crc ", OR, " mcrc ", OR, " colonic "), AND, (" cancer ", OR, " cancers ", OR, " tumor ", OR, " tumors ", OR, " neoplasm ", OR, " neoplasms ", OR, " adenoma ", OR, " adenomas ", OR, " carcinoma ", OR, " carcinomas ", OR, " adenocarcinoma ", OR, " adenocarcinoma ", OR, " malignant ", OR, " malignancy "), AND, (" chemotherapy ", OR, " chemoradiation ", OR, " monoclonal, antibody ", OR, " adjuvant, therapy ", OR, " chemoradiotherapy ", OR, " chemoradiotherapies ", OR, " radiotherapy ", OR, " radiochemotherapies ", OR, " MOAB ", OR, " XELOX ", OR, " fluorouracil ", OR, " irinotecan ", OR, " cetuximab ", OR, " fluoropyrimidine ", OR, " oxaliplatin ", OR, " CPT, 11 ", OR, " 5-FU ", OR, " FOLFOX ", OR, " bevacizumab ", OR, " FOLFIRI ", OR, " XELIRI ", OR, " leucovorin ", OR, " capecitabine ", OR, " FOLFOXIRI ", OR, " CAPOX ", OR, " CAPOXB ", OR, " tamoxifen ", OR, " anti-EGFR, antibody ", OR, " anti-EGFR ", OR, " cisplatin ", OR, " panitumumab ", OR, " platinum ", OR, " tegafur ", OR, " erbitux ", OR, " erlotinib ", OR, " gefitinib ", OR, " tarceva ", OR, " iressa ", OR, " vectibix ", OR, " nimitumumab ", OR, " mazatumumab "), AND, (" response ", OR, " survival ", OR, " outcome ", OR, " resistance ", OR, " outcomes ", OR, " recurrence ", OR, " relapse ", OR, " toxicity ", OR, " responder ", OR, " responders ", OR, " adverse, events ", OR, " adverse, event ", OR, " adverse, effect ", OR, " adverse, effects "), AND, (" polymorphism ", OR, " polymorphisms ", OR, " polymorphic ", OR, " genetic, variant ", OR, " genetic, variants ", OR, " mutation ", OR, " muton ", OR, " saltation ", OR, " break ", OR, " mutations ", OR, " mutons ", OR, " expression ", OR, " express ").

Chinese retrieval formula: ("colorectal cancer", OR, "Cancer") AND, ("Chemotherapy", OR, "Radiation", OR, "Monoclonal antibody", OR, "Treatment", OR, "MOAB" , OR, "XELOX", OR, "FU", OR, "irinotecan", OR, "cetuximab", OR, "oxaliplatin", OR, "platinum", OR, "CPT, 11 ", OR," 5-FU ", OR," FOLFOX ", OR," bevacizumab ", OR," FOLFIRI ", OR," XELIRI ",, OR," capecitabine ", OR," FOLFOXIRI ", OR," CAPOX ", OR," CAPOXB ", OR," tamoxifen ", OR," panitumumab ", OR," tegafur ", OR," Erbitux ", OR," erlotinib ", OR," gefitinib "), AND, (" sensitivity ", OR," survival ", OR," survive ", OR," relapse ", OR," toxic ", OR , "side effects", OR, "prognosis", OR, "forecast", OR, "efficacy", OR, "drug reaction"), AND, ("polymorphism", OR, "mutation", OR, "expression ", OR," micro-satellites ", OR," biomarkers ", OR," molecular markers ").

The list of references of document is included in inspection in, if used identical research object, chooses the research of all the identical research objects sample size maximum or that comprise.

2. formulating document adopts and exclusion standard:

Adopt standard and exclusion standard according to following document, thereby filter out 2325 pieces of standard compliant literature summaries.

Adopt standard: 1. research must be the clinical research of carrying out with artificial research object; 2. research must have the treatment intervention means, except the placebo; 3. research contents must have been explored one or more relevant biomarker biomarkers for the treatment of, comprises gene mutation, gene pleiomorphism, mRNA expression, protein expression level, microRNA and little satellite; 4. clinical efficacy must be one or more in OS, PFS, chemotherapy drug susceptibility, recurrence, the toxic and side effect etc.; 5. only include one of them document of systematic review that type meets randomized controlled trial systematic review, randomized controlled trial, cohort study, cohort study in.

Exclusion standard: the basic medical research that does not 1. have molecule, cell and the animal of direct relation with medical practice; 2. study sample size＜20 examples.

3. the preliminary screening of data analysis and biomarker:

According to standard compliant literature summary, initial analysis literature research type and the patient characteristics, therapeutic scheme, the final result index of measurement effect, the observing time of final result of including research in.According to the research type literature summary is divided into original research and systematic review, and the frequency of occurrences and the regularity of distribution of biomarker gene polymorphic type, sudden change, expression type analysis and colorectal cancer treatment mark of correlation thing, make up the biomarker storehouse.

The screening biomarker meets one of following standard in the storehouse: 1. the systematic review document is arranged in the evidence; 2. randomized controlled trial research document is arranged in the evidence; 3. cohort study's quantity of document 〉=5 piece in the evidence.The selection result is as shown in table 2.

The standard compliant colorectal cancer treatment of table 2 relevant biomarkers thing

4. the grading search of biomarker clinical evidence:

The biomarker that screens is carried out the hierarchical search of evidence, be illustrated in figure 1 as " ERCC1 " as the grading search process of colorectal cancer treatment prognostic markers thing relevant clinical evidence.

Search query, 1 = ("rectal", OR, "Rectum", OR, "Colon", OR, "Colonic", OR, "Colorectal", OR, "Crc", OR, "Mcrc", OR, "Colonic "), AND, (" cancer ", OR," cancers ", OR," tumor ", OR," tumors ", OR," neoplasm ", OR," neoplasms ", OR," adenoma ", OR," adenomas ", OR," carcinoma ", OR," carcinomas ", OR," adenocarcinoma ", OR," adenocarcinoma ", OR," malignant ", OR," malignancy "), AND, (" excision, repair, cross-complementing , rodent, repair, deficiency ", OR," ERCC-1 ", OR," ERCC1 ", OR," ERCC ", OR," excision, repair, cross-complementing, rodent, repair, deficiency,, complementation, group, 1, protein ",), AND, (" response ", OR," survival ", OR," outcome ", OR," resistance ", OR," outcomes ", OR," recurrence ", OR," toxicity ", OR, "relapse");

Retrieval type 2 = ("RCT", OR,, "RCTs", OR, "Randomized, Controlled, Trial", OR, "Randomized-, Controlled, Trial", OR, "Randomized-Controlled-Trial", OR,, "Randomized, Controlled, Trials", OR, "Randomized-Controlled, Trials", OR, "Randomized-Controlled-Trials", OR, "Randomized, Clinical, Trial", OR, "Randomized-Clinical, Trial", OR, "Randomized-Clinical-Trial", OR, "Randomized, Clinical, Trials", OR, "Randomized-Clinical, Trials", OR, "Randomized-Clinical-Trials",);

Retrieval type 3=(" meta-analysis " OR " metaanalysis " OR " meta-analyses " OR " metaanalyses " OR " data pooling " OR " data poolings " OR " meta analysis " OR " meta analyses " OR " meta-analyze " OR " meta analyze " OR " metaanalyze " OR " meta-analytic " OR " meta analytic " OR " metaanalytic " OR " systematic review " OR " systematic reviews " OR " systematic-review " OR " systematic-reviews ");

Retrieval type 4=(" straight colon cancer " OR " intestinal cancer ") AND (" ERCC-1 " OR " ERCC1 " OR " ERCC " OR " excision repair cross-complementing ") AND (" susceptibility " OR " life cycle " OR " existence " OR " recurrence " OR " toxicity " OR " spinoff " OR " prognosis " OR " prediction " OR " curative effect " OR " drug response ");

Retrieval type 5=(" RCT " OR " RCTs " OR " randomized controlled trial " OR " contrast at random " OR " at random ");

Retrieval type 6=(" meta-analysis " OR " meta " OR " systematic review " OR " meta-analysis ").

The remarks explanation:

(1) at English database MEDLINE, EMBASE, Cochrane, TRIP use following search strategy, and be 1980-2012.12.30 retrieval time.：

The retrieval type of the systematic review of randomized controlled trial: retrieval type 1 AND retrieval type 2 AND retrieval types 3;

The retrieval type of randomized controlled trial: retrieval type 1 AND retrieval type 2;

The retrieval type of cohort study's systematic review: retrieval type 1 AND retrieval type 3 NOT (retrieval type 1 AND retrieval type 2 AND retrieval types 3);

The retrieval type of cohort study: retrieval type 1 NOT (retrieval type 1 AND (retrieval type 2 OR retrieval types 3)).

(2) at Chinese database CMDisc, use following search strategy among the CNKI, be 1980-2012.12.30 retrieval time.

The retrieval type of the systematic review of randomized controlled trial: retrieval type 4 AND retrieval types 5 AND retrieval types 6;

The retrieval type of randomized controlled trial: retrieval type 4 AND retrieval types 5;

The retrieval type of cohort study's systematic review: retrieval type 4 AND retrieval types 6 NOT (retrieval type 4 AND retrieval types 5 AND retrieval types 6);

The retrieval type of cohort study: retrieval type 4 NOT (retrieval type 4 AND (retrieval type 5 OR retrieval types 6)).

According to evidence grade (table 1) and evidence 1a＞1b＞2a＞2b right of priority, with the sequential search ERCC1 of the systematic review of randomized controlled trial, randomized controlled trial, systematic review, all the correlative study retrieval types clinical evidence relevant with the colorectal cancer treatment.Result for retrieval shows, about ERCC1 as 0 piece of the systematic review of the randomized controlled trial of oxaliplatin treatment colorectal cancer prognostic markers thing correlative study, 2 pieces of 5 pieces of randomized controlled trials and cohort study's systematic reviews.

Meeting document adopts and 2 pieces of 1 piece of the randomized controlled trial of exclusion standard and cohort study's systematic reviews.

5. the quality evaluation of biomarker clinical evidence:

According to the high-quality evidence judgment criteria of formulating 1 piece of randomized controlled trial and the 2 pieces of systems of cohort study that collect are carried out quality evaluation by 2 valuation officers, and consistently think: ERCC1 gene and result for the treatment of correlation test in the randomized controlled trial do not adopt contrast method at random, a difference patient's pattern detection genotype to 3 groups of different therapeutic schemes, do not compare analysis between group, and sample size in the group＜200 examples, so this evidence does not meet high quality standards.The systematic review of 2 pieces of cohort studies all satisfies: 1. the systematic review research and design has clear and definite search strategy, and searching database comprises MEDLINE and EMBASE, and clear and definite including in and exclusion standard arranged; 2. the original of including in satisfies: sample wide material sources, clear and definite by stages, sample size 〉=300 examples is lost visit rate＜20%, the existence effect follow up a case by regular visits to time＞1 year; 3. statistical method is reasonable, and carries out heterogeneity and detect; 4. prognostic indicators such as amalgamation result PFS, recurrence, toxicity are clear and definite, do not have and to deliver bias (p〉0.05).

According to standard, ERCC1 has as the high-quality evidence of oxaliplatin treatment colorectal cancer prognosis biomarker: 2 pieces of cohort study's systematic reviews.

6. the data analysis of biomarker clinical evidence and statistics:

2 pieces of systematic reviews are all studied ERCC1 gene C 118T polymorphism, but conspicuousness is inconsistent as a result.Be 2008 the 1st piece of systematic review retrieval time, comprises 4 pieces of originals; Be 2011 the 2nd piece of systematic review retrieval time, comprises 9 pieces of originals (wherein 4 pieces of originals are consistent with the 1st piece of systematic review) and 968 routine colorectal cancer patients, simultaneously the final result index adopted the meta analysis.According to selecting more, the more rational standard of test design of sample, adopt the result of the 2nd piece of systematic review at last.

7. colorectal cancer treatment prognostic markers thing screening:

According to screening criteria, screening obtains ERCC1 gene C 118T polymorphism and is one and for the Primary Care prognosis the relevant biomarker with potential using value is arranged with the oxaliplatin with colorectal cancer patients.Evidence shows, carry the allelic colorectal cancer patients of ERCC1 gene 11 8T to the oxaliplatin drug effect present conspicuousness the low reaction rate (OR=0.53,95%CI=0.35-0.81).The probability of carrying allele T patient tumors progress simultaneously increases tool significance,statistical (HR=1.69; 95%CI=1.05-2.70), and reduced total survival rate (HR=2.03 of patient; 95%CI=1.60-2.59).In addition, a plurality of colorectal cancer treatment prognosis biomarkers (table 3) have also been screened by this method.

Table 3 colorectal cancer treatment prognosis biomarker The selection result

8, result verification:

2012, the U.S. state-run comprehensive cancer network (National Comprehensive Cancer Network, NCCN) guide is recommended, with KRAS, BRAF, these 4 labels of MSI, UGT1A1*28 prognosis biomarker that treatment is correlated with as colorectal cancer, above label all can obtain by the screening technique with this patent, as table 3.

9, demonstration test:

The biomarker ERCC1 gene 11 8 C＞T polymorphism that filters out for further checking the present invention is as the accuracy for the treatment of prognostic markers thing, and retrospective including in accepted oxaliplatin treatment and colorectal cancer patients 100 examples of complete prognosis record are arranged.ERCC1 gene 11 8 C＞T polymorphism to the tissue specimen of colorectal cancer patients detects the PCR-RFLP method that adopts, PCR expansion primer and restriction enzyme such as table 4.The result shows, carries the homozygotic colorectal cancer patients of ERCC1 allele 118 C/C to oxaliplatin drug effect reaction low (OR=0.42,95% CI=0.21-0.83), and the result is remarkable.Simultaneously, carry ERCC1 allele C/C homozygote patient accept after the oxaliplatin treatment significant prolongation 5 years total life cycles (HR=0.32,95%CI=0.13-0.79).Therefore, the ERCC1 gene 11 8 C＞T polymorphism that obtains with the screening of this patent method can be used as the prognosis biomarker that colorectal cancer patients is accepted oxaliplatin treatment.

PCR expansion primer and the restriction enzyme of table 4 PCR-RFLP detection method

Contrast test: can not be as treatment prognostic markers thing for biomarker IL8 gene 251 T＞A polymorphism that checking the present invention gets rid of, retrospective in the same way including in accepted the 5 FU 5 fluorouracil treatment and colorectal cancer patients 100 examples of complete prognosis record arranged.IL8 gene 251 T＞A polymorphism to the patient tissue sample detects the PCR-RFLP method that adopts, PCR expansion primer and restriction enzyme such as table 4.The result shows, the colorectal cancer patients of carrying IL8 allele T to the 5 FU 5 fluorouracil drug effect reaction (OR=1.64,95%CI=0.96-2.8), but the result is not remarkable than carrying allele A patient height.Compare with carrying T/T homozygote patient, carry T/A heterozygote and T/T homozygote patient 5 years total life cycle of HR and be respectively 1.19 (95%CI=0.73-1.94) and 1.03 (95%CI=0.57-1.85).Therefore, patient's drug effect reaction and 5 years total life cycles and IL8 gene 251 T＞A polymorphism do not have significant correlation, and biomarker IL8 gene 251 T＞A polymorphism that the present invention gets rid of can not be accepted the prognosis biomarker of 5 FU 5 fluorouracil treatment as colorectal cancer patients.

At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.

Claims (4)

1. colorectal cancer disease is treated the screening technique of prognosis biomarker, it is characterized in that may further comprise the steps:

1), formulate the search strategy of preliminary screening biomarker:

According to PICOS principle deterministic retrieval word, and the descriptor index of employing cloth woods connector formula;

Utilize term to comprise MEDLINE at database, EMBASE, Cochrane, TRIP, Wei Pu (CPVIP), Chinese biological medical journals bibliographic data base (CMDisc), comprehensive searching document in database and the total storehouse of Chinese knowledge resource (CNKI) incomparably; The list of references of document is included in inspection in, if used identical research object, chooses the research of all the identical research objects sample size maximum or that comprise;

2), formulating document adopts and exclusion standard:

Adopt and exclusion standard according to document, screen standard compliant literature summary;

Adopt standard: 1. research must be the clinical research of carrying out with artificial research object; 2. research must have the treatment intervention means; 3. research contents must have been explored one or more relevant biomarkers for the treatment of; 4. clinical efficacy must be objective life cycle, drug response rate or bad reaction; 5. only include perspective and retrospective study in;

Exclusion standard: the basic medical research that does not 1. have molecule, cell and the animal of direct relation with medical practice; 2. study sample size＜20 examples;

3), the preliminary screening of data analysis and biomarker:

According to standard compliant literature summary, initial analysis literature research type and the patient characteristics, therapeutic scheme, the final result index of measurement effect, the observing time of final result of including research in, analyze the biomarker regularity of distribution of different document quality, different treatment means, different treatment results, different research types, make up the biomarker storehouse;

The screening biomarker meets one of following standard in the storehouse: 1. the systematic review document is arranged in the evidence; 2. randomized controlled trial research document is arranged in the evidence; 3. cohort study's quantity of document 〉=5 piece in the evidence;

4), formulate the rank of evidence:

According to the evidence classification of colorectal cancer treatment prognosis biomarker, that the evidence classification is as follows:

The requirement of evidence rank evidence

1 a comprises the systematic review of a plurality of homogeneity randomized controlled trials;

At least one randomized controlled trial of 1 b;

2 a comprise the systematic review of a plurality of cohort researchs;

At least one cohort study of 2 b;

5), formulate high-quality evidence judgment criteria:

4 grades of different literature research types are formulated the judgment criteria of high-quality evidence with reference to Newcastle-Ottawa Scale standard;

6), the grading search of biomarker clinical evidence and quality evaluation:

Evidence rank 1a＞1b＞2a＞2b is comply with in the retrieval of biomarker clinical evidence, with the biomarker relevant clinical evidence in the sequential search primary dcreening operation of the systematic review of randomized controlled trial, randomized controlled trial, systematic review, all correlative study retrieval types, include in and meet document and adopt clinical evidence with exclusion standard, according to high-quality evidence judgment criteria the clinical evidence of including in is carried out quality evaluation again;

7), the data analysis of biomarker clinical evidence and statistics:

Only the high-quality clinical evidence of biomarker is carried out data analysis and statistics, as evidence 1a, when having only one piece of systematic review in the 2a class, this evidence directly is included into;

When many pieces of systematic reviews occurring: (1) includes all systematic reviews in if the result is significantly consistent or unanimity is not remarkable; When (2) result is inconsistent, need to consider that single piece of systematic review includes differences such as the scheme of the original scope of research and quantity, test design, study population's feature, statistical method in, the systematic review of selecting to include in satisfies standard: 1. sample is more; 2. test design is more reasonable;

Evidence 1b, when having only one piece of systematic review among the 2b, this evidence directly is included into; When many pieces of researchs occurring, use meta to analyze amalgamation result, Q-test estimates heterogeneous, and searches heterogeneous source with the subgroup analysis, delivers bias with Begg ' s funnel figure check; The amalgamation result index is included the evidence storehouse in;

8), treatment prognostic markers thing screening:

Prognostic indicators such as The selection result OS, PFS, recurrence, toxicity significantly (p＜0.05) from the evidence of including in do not have the biomarker of delivering bias (p〉0.05).

2. colorectal cancer disease according to claim 1 is treated the screening technique of prognosis biomarker, and it is characterized in that: described step 5) comprises the steps:

(1) systematic review of randomized controlled trial satisfies: 1. the systematic review research and design has clear and definite search strategy, and searching database comprises MEDLINE and EMBASE; The clear and definite standard of including in is arranged; 2. the original of including in satisfies: diagnosing tumor is clear and definite by stages, and sample size 〉=50 examples is lost visit rate＜20%, the existence effect follow up a case by regular visits to time＞2 year, the research and design method is clear and definite, research mode adopts single blind or double blinding; 3. statistical method is reasonable, and carries out heterogeneity and detect; 4. prognostic indicators such as amalgamation result PFS, recurrence, toxicity are clear and definite, do not have and to deliver bias (p〉0.05);

(2) randomized controlled trial satisfies: 1. diagnosing tumor is clear and definite by stages, sample size 〉=200 examples; 2. lose visit rate＜20%, the existence effect follow up a case by regular visits to time＞2 year; 3. research mode adopts single blind or double blinding; 4. the biomarker object detecting method is clear and definite; 5. prognosis final result indexs such as PFS, recurrence, toxicity are clear and definite; 6. positive findings threshold value definition clear-cut;

(3) systematic review of cohort study's test satisfies: 1. the systematic review research and design has clear and definite search strategy, and searching database comprises MEDLINE and EMBASE; Clear and definite including in and exclusion standard arranged; 2. the original of including in satisfies: sample wide material sources, clear and definite by stages, sample size 〉=300 examples is lost visit rate＜20%, the existence effect follow up a case by regular visits to time＞1 year; 3. statistical method is reasonable, and carries out heterogeneity and detect; 4. prognostic indicators such as amalgamation result PFS, recurrence, toxicity are clear and definite, do not have and to deliver bias (p〉0.05);

(4) cohort study test is satisfied: 1. sample wide material sources, sample size 〉=300 examples; 2. diagnosing tumor is clear and definite by stages; 3. lose visit rate＜20%, the existence effect follow up a case by regular visits to time＞2 year; 4. the research and design method is reasonable, and the biomarker object detecting method is clear and definite; 5. prognosis final result indexs such as PFS, recurrence, toxicity are clear and definite; 6. positive findings threshold value definition clear-cut; 7. deliver periodical IF＞2.

3. colorectal cancer disease according to claim 2 is treated the screening technique of prognosis biomarker, it is characterized in that: in the described step 1):

foreign language database retrieval formula: (" rectal ", OR, " rectum ", OR, " colon ", OR, " colonic ", OR, " colorectal ", OR, " crc ", OR, " mcrc ", OR, " colonic "), AND, (" cancer ", OR, " cancers ", OR, " tumor ", OR, " tumors ", OR, " neoplasm ", OR, " neoplasms ", OR, " adenoma ", OR, " adenomas ", OR, " carcinoma ", OR, " carcinomas ", OR, " adenocarcinoma ", OR, " adenocarcinoma ", OR, " malignant ", OR, " malignancy "), AND, (" chemotherapy ", OR, " chemoradiation ", OR, " monoclonal, antibody ", OR, " adjuvant, therapy ", OR, " chemoradiotherapy ", OR, " chemoradiotherapies ", OR, " radiotherapy ", OR, " radiochemotherapies ", OR, " MOAB ", OR, " XELOX ", OR, " fluorouracil ", OR, " irinotecan ", OR, " cetuximab ", OR, " fluoropyrimidine ", OR, " oxaliplatin ", OR, " CPT, 11 ", OR, " 5-FU ", OR, " FOLFOX ", OR, " bevacizumab ", OR, " FOLFIRI ", OR, " XELIRI ", OR, " leucovorin ", OR, " capecitabine ", OR, " FOLFOXIRI ", OR, " CAPOX ", OR, " CAPOXB ", OR, " tamoxifen ", OR, " anti-EGFR, antibody ", OR, " anti-EGFR ", OR, " cisplatin ", OR, " panitumumab ", OR, " platinum ", OR, " tegafur ", OR, " erbitux ", OR, " erlotinib ", OR, " gefitinib ", OR, " tarceva ", OR, " iressa ", OR, " vectibix ", OR, " nimitumumab ", OR, " mazatumumab "), AND, (" response ", OR, " survival ", OR, " outcome ", OR, " resistance ", OR, " outcomes ", OR, " recurrence ", OR, " relapse ", OR, " toxicity ", OR, " responder ", OR, " responders ", OR, " adverse, events ", OR, " adverse, event ", OR, " adverse, effect ", OR, " adverse, effects "), AND, (" polymorphism ", OR, " polymorphisms ", OR, " polymorphic ", OR, " genetic, variant ", OR, " genetic, variants ", OR, " mutation ", OR, " muton ", OR, " saltation ", OR, " break ", OR, " mutations ", OR, " mutons ", OR, " expression ", OR, " express ")

Chinese retrieval formula: ("colorectal cancer", OR, "Cancer") AND, ("Chemotherapy", OR, "Radiation", OR, "Monoclonal antibody", OR, "Treatment", OR, "MOAB" , OR, "XELOX", OR, "FU", OR, "irinotecan", OR, "cetuximab", OR, "oxaliplatin", OR, "platinum", OR, "CPT, 11 ", OR," 5-FU ", OR," FOLFOX ", OR," bevacizumab ", OR," FOLFIRI ", OR," XELIRI ",, OR," capecitabine ", OR," FOLFOXIRI ", OR," CAPOX ", OR," CAPOXB ", OR," tamoxifen ", OR," panitumumab ", OR," tegafur ", OR," Erbitux ", OR," erlotinib ", OR," gefitinib "), AND, (" sensitivity ", OR," survival ", OR," survive ", OR," relapse ", OR," toxic ", OR , "side effects", OR, "prognosis", OR, "forecast", OR, "efficacy", OR, "drug reaction"), AND, ("polymorphism", OR, "mutation", OR, "expression ", OR," micro-satellites ", OR," biomarkers ", OR," molecular markers ");

The list of references of document is included in inspection in, if used identical research object, chooses the research of all the identical research objects sample size maximum or that comprise.

4. utilize in the claim 1 ~ 3 any one method screening and colorectal cancer disease treatment prognosis biomarker, it is characterized in that for following any one:

KRAS、TYMS、TYMS、EGFR、UGT1A1*28、DPYD、PIK3CA、ERCC1、PTEN、VEGFR、BRAF、GST?P1、XRCC1、MTHFR、MSI、ERCC2。

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