【作者】 苏彤；

【导师】 曹广文；

【作者基本信息】 第二军医大学 ， 流行病学与卫生统计学， 2012， 硕士

【摘要】 【研究目的】肺癌是我国肿瘤死亡的第一位死因，目前手术和化疗的效果均不理想，关键是要根据患者的遗传背景进行个体化治疗。本研究目的是探讨DNA修复系统的五个重要基因包括X射线交叉互补基因1（x-ray repair cross-complementing group1,XRCC1）、切除修复交叉互补基因1（excision repair cross-complementing group1,ERCC1）、着色性干皮病基因D（xeroderma pigmentosum group D, XPD）、X射线交叉互补基因3（x-ray repair cross-complementing group3, XRCC3）、乳腺癌易感基因1（breast cancer susceptibility gene1, BRCA1）和多药耐药基因（multiple drug resistancegene1，MDR1）的单核苷酸多态性（single nucleotide polymorphisms, SNPs）分别与肺癌早期手术患者和晚期化疗患者治疗效果和生存时间的关系；探讨蛋白酪氨酸磷酸酶受体D基因（protein-tyrosine phosphatase receptor delta gene, PTPRD）多态性与肺癌易感性的关系，在此基础上进一步明确PTPRD SNP与肺癌预后的关系，从而为肺癌早期发现和预后预测提供依据。【研究方法】共收集377例肺癌患者和754例健康对照，提取外周血DNA，采用Taqman探针法检测XRCC1Arg399Gln (rs25487)，ERCC1Asn118Asn (rs11615)，XPDLys751Gln (rs13181)，XRCC3A4541G (rs1799794)，BRCA1Ser1613Gly (rs1799966)，MDR1Ile1145Ile (rs1045642)和PTPRD G4254C (rs2279776)。对肺癌患者进行临床疗效评价和预后随访。以肿瘤完全缓解（CR）、部分缓解（PR）、病情稳定（SD）为治疗受益，以肿瘤增大或病情进展（PD）为非受益。采用χ2检验和非条件Logistic回归模型确定SNPs与肺癌易感性和化疗效果的关系，采用Kaplan-Meier方法Log-rank检验和Cox比例风险回归模型确定SNPs与肺癌患者生存时间的关系。【结果】1. BRCA1Ser1613Gly和MDR1Ile1145Ile与晚期非小细胞肺癌（non-small celllung cancer, NSCLC）化疗效果显著相关，BRCA1G等位基因（AG+GG）和MDR1T等位基因（CT+TT）的化疗受益率均高于野生型（P=0.013，0.045）。ERCC1Asn118Asn、XPD Lys751Gln、XRCC1Arg399Gln和XRCC3A4541G与化疗效果无关。这6个SNPs存在一定联合作用，携带变异等位基因（XRCC1A, ERCC1T, XPD C, XRCC3G,BRCA1G或MDR1T）越多，化疗受益率越高(P=0.024)。2. BRCA1Ser1613Gly是晚期NSCLC化疗患者生存时间的独立影响因素，BRCA1AG+GG比AA型生存时间更长(Hazard Ratio [HR]=0.617,95%CI=0.402-0.948,P=0.028)。按临床特征对晚期NSCLC化疗患者分层分析显示，在鳞癌、吸烟、PS评分=1和Ⅲ期化疗患者中BRCA1AG+GG生存时间比AA型更长（Log-rank P<0.05），在腺癌患者中XRCC1GA+AA生存时间比GG型更长（Log-rank P=0.038，HR=0.496，95%CI=0.246-0.997）。3. ERCC1Asn118Asn是早期NSCLC手术患者生存时间的独立影响因素，ERCC1TT型比CC+CT型生存时间短（HR=3.087，95%CI=1.197-7.961，P=0.020）。对早期NSCLC手术患者分层分析显示，在Ⅲ期手术患者和鳞癌手术患者中XPD AC型的生存时间明显少于AA型（Log-rank P=0.016，0.008）；在Ⅲ期手术患者和接受辅助化疗的手术患者中XRCC1GA+AA型的生存时间明显长于GG型（Log-rank P=0.004，0.050）。4. PTPRD G4254C与肺癌易感性相关，GC+CC型比GG型患肺癌的风险更高（P=0.039, odds ratio [OR]=1.298,95%CI=1.013-1.663），在吸烟者中关系更为显著（校正OR=1.563，95%CI=1.013-2.412）。PTPRD G4254C与肺癌化疗效果和总体生存时间之间无明显关联。对肺癌早期手术患者分层分析显示，在女性手术患者、腺癌手术患者和I期手术患者中PTPRD CC型生存时间明显少于GG+GC型（Log-rank P=0.014，0.009，0.000）。【结论】 BRCA1Ser1613Gly和MDR1Ile1145Ile与晚期NSCLC化疗效果显著相关，联合分析多个DNA修复基因和MDR1基因的SNPs可能对指导化疗药物的选择更有帮助。BRCA1Ser1613Gly和ERCC1Asn118Asn分别与晚期NSCLC化疗患者和早期NSCLC手术患者生存时间明显相关，可作为肺癌预后预测指标。PTPRD G4254C与肺癌易感性相关，但与化疗效果和生存时间的关系不明显。更多还原

【Abstract】 [Objective] Lung cancer is the first leading cause of cancer-related death in China.Currently, the outcome of resection or chemotherapy for lung cancer is not satisfactory.Individualized treatment according to specific genetic background is very important forimproving outcome. One aim of this study is to investigate the association of singlenucleotide polymorphism (SNP) of a group of important genes including x-ray repaircross-complementing group1(XRCC1), excision repair cross-complementing group1(ERCC1), xeroderma pigmentosum group D (XPD), x-ray repair cross-complementinggroup3(XRCC3), breast cancer susceptibility gene1(BRCA1) and multiple drugresistance gene1(MDR1) polymorphisms with therapeutic results of the chemotherapyand survivals of lung cancer patients. We also investigated the association ofprotein-tyrosine phosphatase receptor delta gene (PTPRD) polymorphism with thesusceptivity and the prognosis of lung cancer in Chinese population, providing evidencefor early detection and prognosis prediction of this malignancy.[Methods] A total of377patients with lung cancer and754healthy controls wereenrolled in the study. DNA was isolated from peripheral bloods of the study subjects.Seven polymorphisms (XRCC1Arg399Gln，ERCC1Asn118Asn，XPD Lys751Gln，XRCC3A4541G，BRCA1Ser1613Gly，MDR1Ile1145Ile and PTPRD G4254C) weregenotyped by TaqMan method. The chemotherapeutic effect and overall survival (OS) inpatients with lung cancer were evaluated. Patients with complete response (CR), partialresponse (PR) and stable disease (SD) were defined as patients with clinical benefit, andpatients with progressive disease (PD) were defined as patients without clinical benefit.Associations between polymorphisms and lung cancer risk or chemotherapeutic effectwere estimated using χ2test and unconditional logistic regression model. Associationsbetween polymorphisms and OS were estimated using Kaplan-Meier methods andLog-rank test for univariate analysis and Cox proportional hazards model for multivariateanalysis.[Results]1. BRCA1Ser1613Gly and MDR1Ile1145Ile significantly correlated withchemotherapeutic effect in advanced non-small cell lung cancer (NSCLC) patients. TheBRCA1G allele carriers (AG+GG) and the MDR1T allele carriers (CT+TT) had betterclinical benefit than wild type carriers (P=0.013, P=0.045, respectively). No significantassociations were found between ERCC1Asn118Asn, XPD Lys751Gln, XRCC1 Arg399Gln or XRCC3A4541G and clinical benefit. Furthermore, we found that the6polymorphisms had interaction in the drug response, a greater number of variant alleles(XRCC1A, ERCC1T, XPD C, XRCC3G, BRCA1G or MDR1T) was associated withbetter clinical benefit (P=0.024).2. Advanced NSCLC patients treated with chemotherapy with BRCA1G allele (AG+GG)had a significantly better OS compared with advanced NSCLC patients with AA genotype(Hazard Ratio [HR]=0.617,95%CI=0.402-0.948, P=0.028). Stratified advanced NSCLCpatients according to clinical characters, BRCA1AG+GG genotype was significantlyassociated with better OS among advanced NSCLC patients with squamous cell carcinoma,smoking habit, performance status1, and stage III (Log-rank P<0.05); XRCC1GA+AAgenotype was significantly associated with better OS among advanced NSCLC patientswith adenocarcinoma (Log-rank P=0.038，HR=0.496，95%CI=0.246-0.997).3. Postoperative early-stage NSCLC patients with ERCC1TT genotype had a significantlyworse OS compared with early-stage NSCLC patients with CC+CT genotype (HR=3.087，95%CI=1.197-7.961，P=0.020). Stratified early-stage NSCLC patients according toclinical characters, XPD AC genotype was significantly associated with worse OS amongearly-stage NSCLC patients with stage III and squamous cell carcinoma (Log-rankP=0.016,0.008, respectively); XRCC1GA+AA genotype was significantly associated withbetter OS among early-stage NSCLC patients with stage III and adjuvant chemotherapy(Log-rank P=0.004,0.050, respectively).4. PTPRD G4254C was significantly associated with the risk of lung cancer, GC+CCgenotype had a significantly increased lung cancer risk compared with GG genotype(P=0.039, OR=1.298,95%CI=1.013-1.663). Similar association existed in smokers(ajusted OR=1.563，95%CI=1.013-2.412). In total, no significant associations were foundamong PTPRD G4254C and both chemotherapeutic effect and OS. Stratified postoperativeearly-stage NSCLC patients according to clinical characters, PTPRD CC genotype wassignificantly associated with worse OS among female patients, adenocarcinoma patientsand stage I patients (Log-rank P=0.014,0.009,0.000, respectively).[Conclusion] BRCA1Ser1613Gly and MDR1Ile1145Ile may affect the response ofadvanced NSCLC patients to chemotherapy. It can be useful to research combinations ofSNPs for customizing chemotherapy. BRCA1Ser1613Gly may be important prognosticmarker in advanced NSCLC patients treated with chemotherapy, and ERCC1Asn118Asnmay be important prognostic marker in early-stage NSCLC patients treated with operation. PTPRD G4254C polymorphism is significantly associated with a higher risk of developinglung cancer, but does not significantly affect chemotherapeutic effect and OS in lungcancer patients.更多还原