【作者】 王莹；

【导师】 吴长利；

【作者基本信息】 天津医科大学 ， 外科学， 2013， 硕士

【摘要】 目的在细胞周期蛋白E1基因(CCNE1)的5’非编码区存在多态性位点rs8102137,在受体相互作用蛋白2基因(RIP2)内含子区存在多态性位点rs42490,目前这两个多态位点与肿瘤发生及发展关系尚不明确,本文意在探讨这两位点的多态性与膀胱癌的发病风险及膀胱癌的病理分级分期之间的关系。方法本研究课题获得天津医科大学医学伦理委员会批准。共包含386例研究对象。实验组为膀胱移行细胞癌患者,无其他肿瘤病史,共176例；对照组为随即健康体检者共210例。膀胱癌组根据肿瘤细胞的分化程度,按照WHO2004分级法分级,分为：低度恶性倾向尿路上皮乳头状瘤(Papillary urothelial neoplasms of low malignant potential, PUNLMP)、乳头状尿路上皮癌,低分级、乳头状尿路上皮癌,高分级三个级别。按照国际抗癌联盟(UICC)2009年第7版TMN分期法对膀胱癌进行分期,分为Tis原位癌、Ta非浸润性乳头状癌、T1肿瘤侵及上皮下结缔组织、T2肿瘤侵犯肌层、T3肿瘤侵犯膀胱周围组织、T4侵犯以下任一器官或组织,如前列腺、子宫、阴道、盆壁和腹壁。非浸润性膀胱癌包括Tis、Ta、T1,浸润性膀胱癌包括T2、T3、T4。对照组将有明确膀胱癌或肿瘤家族史、腺性增生性膀胱炎、前列腺特异抗原(PSA)大于4.0ng/ml的前列腺增生患者、甲胎蛋白(AFP)大于400mg/ml、癌胚抗原(CEA)大于15.0ug/L等患者排除。每例患者留取血标本5ml,应用EDTA抗凝,分别提取DNA,通过聚合酶链反应(PCR)—测序方法来检测细胞周期蛋白E1基因(CCNE1) rs8102137和受体相互作用蛋白2基因(RIP2) rs42490的多态性。通过经尿道膀胱肿瘤电切术、膀胱部分切除术及膀胱癌根治性切除术确定膀胱癌患者的诊断、病理分级及分期。采用SPSS13.0for Windows统计软件包,以χ2检验检测对照组中基因型分布是否符合Hardy-Weinberg平衡,分析病例组和对照组中等位基因和基因型的分布差异。对病例组与对照组间基因型的分布情况进行比较,并应用χ2检验和SPSS13.0for Windows统计软件分析CCNE1(rs8102137)和RIP2(rs42490)各基因型与膀胱癌遗传易感性及其病理分级、分期的关系。检验水平a=0.05,以比值比(OR)和95%的可信区间(Confidence Intervals, CI)表示相对危险度。结果1. CCNE1和RIP2基因吻合度检测对照组中CCNE1(rs8102137)和RIP2(rs42490)的基因型频率与Hardy-Weinberg平衡相符(P>0.05),说明对照组样本的基因型分布具有良好的群体代表性。2. CCNE1单核苷酸多态性分布特点与膀胱癌遗传易感性的关联检测到CCNE1(rs8102137)的基因型有三种：野生型(T/T)、突变杂合子(T/C)、突变纯合子(C/C)。他们在两组中所占的比例分别为：膀胱癌组59.09%(104/176)、39.20%(69/176)、1.70%(3/176)；对照组69.05%(145/210)、29.52%(62/210)、1.43%(3/210)。通过比较CCNE1(rs8102137)各基因型在两组之间的分布比例发现,含稀有等位基因C的突变基因型(T/C+C/C)在膀胱癌组和对照组中存在明显的分布差异(P<0.05)。膀胱癌组CCNE1突变基因型(T/C+C/C)的频率(40.91%)显著高于对照组(30.95%)(P<0.05, OR=1.54,95%CI1.02-2.45)。3.RIP2单核苷酸多态性分布特点与膀胱癌遗传易感性的关联检测到RIP2(rs42490)的基因型有三种：野生型(G/G)、突变杂合子(G/A)、突变纯合子(A/A)。他们在两组中所占的比例分别为：膀胱癌组27.27%(48/176)、46.02%(81/176)、26.70%(47/176)；对照组37.62%(79/210)、45.71%(96/210)、16.67%(35/210)。通过比较RIP2(rs42490)各基因型在两组间的分布比例发现,含稀有等位基因A的突变基因型(G/A+A/A)在膀胱癌组和对照组中存在明显的分布差异(P<0.05)。膀胱癌组RIP2突变基因型(G/A+A/A)频率(72.73%)显著高于对照组(62.38%)(P<0.05, OR=1.61,95%CI1.04-2.48)。4. CCNE1和RIP2基因型与膀胱癌病理分级、分期的关系CCNE1(rs8102137)和RIP2(rs42490)基因型与膀胱癌病理分级、分期均无显著性相关。结论1. CCNE1(rs8102137)基因型与膀胱癌关系密切,携带稀有等位基因C的个体患膀胱癌的风险显著高于野生型个体；RIP2(rs42490)基因型与膀胱癌关系密切,携带稀有等位基因A的个体患膀胱癌的风险显著高于野生型个体。2. CCNE1(rs8102137)和RIP2(rs42490)基因多态性与膀肌癌的病理分级和临床分期均无显著相关性。更多还原

【Abstract】 ObjectiveAlleles at single nucleotide polymorphisms (SNPs) rs8102137and rs42490within the5’-UTR of CCNE1and the intron of RIP2, respectively, had an uncertain impact on cancer risk. Here, we want to identify the relationship of these two SNPs with Bladder Transitional Cell Carcinoma risk.Materials and MethodsThe study included176patients with BTCC (Bladder Transitional Cell Carcinoma), The control group comprised210healthy blood donors who had a medical examination. A total of5ml of peripheral venous blood was drawn with EDTA as an anticoagulant. And total genomic DNA was isolated from peripheral blood. Detection of genotype variants was analyzed by the polymerase chain reaction-restriction sequencing method. The diagnoses, pathological grade and stage of bladder cancer were all determined according to the pathological reports of transurethral bladder cancer resection, partial cystectomy and radical cystectomy. Hardy-Weinberg equilibrium was checked in control by the goodness of fit χ2test. Genotype and allele frequencies of the CCNE1and RIP2gene in the healthy group and in the bladder cancer group were analyzed using the χ2test. The relationship between polymorphisms and the risk to bladder cancer were analyzed using the odds ratio (OR) and95%confidence intervals (CI). Statistical significance was set at P<0.05. The SPSS version13.0software package was used for analyzes.Results1. The detection of CCNE1and RIP2genetic equilibrium The frequencies of rs8102103and rs42490locus conform to Hardy-Weinberg equilibrium, insuring the reliability of their application to evaluate larger groups.2. Association between CCNEl(rs8102137) genotypes or alleles andsusceptibility of bladder cancerIn patients, the frequencies of the T/T, T/C and C/C genotypes were59.09%(104/176).39.20%(69/176)、1.70%(3/176), respectively. In controls, the frequencies of the T/T, T/C and C/C genotypes were69.05%(145/210)、 29.52%(62/210)、1.43%(3/210), respectively. The frequency of CCNE1(rs8102137) variant allele was significantly higher in patients (40.91%) than in controls (30.95%)(P<0.05, OR=1.54,95%CI1.02-2.45).3. Association between RIP2(rs42490) genotypes or alleles and susceptibility of bladder cancerIn patients, the frequencies of the T/T, T/C and C/C genotypes were27.27%(48/176)、46.02%(81/176)、26.70%(47/176), respectively. In controls, the frequencies of the T/T, T/C and C/C genotypes were37.62%(79/210)、45.71%(96/210)、16.67%(35/210), respectively. The frequency of CCNE1(rs8102137) variant allele was significantly higher inpatients (72.73%) than in controls (62.38%)(P<0.05, OR=1.61,95%CI1.04-2.48).4. CCNE1and RIP2genotypes and pathological grade and clinical stage of bladder cancerFor the CCNE1(rs8102137) and RIP2(rs42490) genotypes, there was no association between the CCNE1(rs8102137) and RIP2(rs42490) Polymorphisms and Pathological grade and clinical stage of bladder cancer. We did not found significant association between the studied polymorphisms with neither tumor grade nor tumor stage.Conclusions1. The genotype T/C and C/C in CCNE1rs8102137showed an increased cancer risk when compared with T carriers in our study population; The genotype G/A and A/A in RIP2rs42490showed an increased cancer risk when compared with G carriers in our study population.2. No significant association between the studied polymorphisms with neither tumor grade nor tumor stage.更多还原