【作者】 王晓明；

【导师】 王建业；

【作者基本信息】 北京协和医学院 ， 泌尿外科（专业学位）， 2014， 博士

【摘要】 第一部分雄激素受体基因(CAG)n重复多态性与中国北方人群前列腺癌患病风险相关性的病例对照研究目的研究雄激素受体(AR)基因(CAG)n重复多态性与中国北方人群前列腺癌(PCa)患病风险的关系。材料和方法回顾性分析中国北方地区PCa患者140例和按年龄匹配的健康男性206例,记录受试者临床信息并用盐析法提取外周血基因组DNA。利用荧光标记引物PCR扩增AR基因第一外显子,毛细管电泳法进行基因分型,然后用Sanger法直接测序验证分型结果。根据基因分型结果推算(CAG)n重复次数。比较AR基因(CAG)。重复次数在PCa组和对照组的分布差异,并分析(CAG)n重复多态性与PCa患者年龄、体重指数(BMI)、前列腺特异抗原(PSA、Gleason评分和临床病理分期的关系。PCa组和对照组间均数比较采用独立样本t检验或秩和检验,两组基因型频率分布差异的比较采用Z检验,计算比值比(OR)及其95%置信区间(95%CI),P<0.05认为具有统计学意义。统计学分析采用SPSS19.0软件完成。结果最终纳入具有完整临床资料和基因分型结果的研究对象共323例,PCa组126例,对照组197例。两组受试者的年龄和BMI无显著性差异(P>0.05)。PCa组AR基因(CAG)n平均重复次数为19.81±3.30,对照组(CAG)。平均重复次数为20.58±2.75,PCa组(CAG)。平均重复次数明显少于对照组(P<0.05)。短(CAG)。重复(<20次)序列AR基因的分布频率在PCa组明显高于对照组(OR=1.70,95%CI:1.06,2.71,P<0.05)；且短(CAG)。重复序列是发生高级别(Gleason评分>7分)PCa的危险因素(OR=4.12,95%CI:1.84,9.23, P<0.01)。AR基因(CAG)。重复多态性在不同年龄、BMI、PSA和临床病理分期的PCa患者中分布无显著性差异(P>0.05)。结论AR基因(CAG)n重复多态性与中国北方人群PCa患病风险相关；短(CAG)。重复(<20次)序列会增加PCa患病风险,且与高级别PCa的发生具有一定相关性。第二部分雄激素受体基因(CAG)n重复多态性与多人群前列腺癌患病风险相关性的Meta分析目的用Meta分析的方法研究雄激素受体基因(CAG)n重复多态性与多人群前列腺癌患病风险的相关性。材料和方法检索2013年8月之前国内外大型数据库中关于AR基因(CAG)n重复多态性与PCa患病风险关系研究的文献。制定文献纳入和排除标准进行筛选,并加入第一部分研究数据。采取双人盲法提取文献数据,用Stata12.0软件Meta分析模块完成分析。首先采用Q检验分析各研究之间的异质性,进行亚组分析挖掘异质性来源。然后采用Mantel-Haenszel固定效应模型或随机效应模型合并效应量分析,计算OR值和95%CI评价AR基因(CAG)n重复多态性与PCa患病风险的相关性,进而按照研究人群种族进行亚组分析。并进行敏感性分析和累积分析评价Meta分析结果的稳定性。最后采用Begg’s检验和Egger’s检验及绘制漏斗图的方法评估潜在发表偏倚,采用剪补法识别和处理潜在发表偏倚。结果检索到相关文献215篇,入选31篇加上我们的研究进行Meta分析。研究人群包括高加索人、亚洲人、非洲人和西班牙人,其中PCa组5,516例、健康对照组6,834例。各研究间存在明显异质性(I2=76.2%,P<0.05),其中以非洲人(I2=88.0%)和亚洲人(I2=81.5%)为研究对象的研究间异质性较大。采用随机效应模型进行合并效应量分析显示,短(CAG)n重复序列AR基因显著增加PCa患病风险(OR=1.61,95%CI：1.34,1.94)。亚组分析提示,短(CAG)。重复序列与高加索人(OR=1.37,95%CI：1.15,1.63)和亚洲人(OR=2.10,95%CI:1.38,3.19) PCa患病风险增加具有显著相关性,而与非洲人和西班牙人PCa患病风险的相关性不明显。敏感性分析和累计分析提示本项Meta分析结果较为稳定。尽管Begg’s检验和Egger’s检验均提示存在显著发表偏倚(P<0.05),但用剪补法剪补前后的合并效应量OR值无显著性差异。结论AR基因(CAG)n重复多态性与PCa患病风险的关系受到人群种族的影响,短(CAG)n重复序列与高加索人和亚洲人PCa患病风险增加呈正相关性；但与非洲人和西班牙人PCa患病风险无明显相关性。更多还原

【Abstract】 Section1Association between androgen receptor gene (CAG)n repeat polymorphisms and the risk of prostate cancer among northern Chinese men:A case-control studyObjectives To derive an association between androgen receptor (AR) gene (CAG) n repeat polymorphisms and prostate cancer (PCa) risk among northern Chinese men.Materials and methods140PCa patients and206age-matched healthy men were analysed retrospectively.Clinical records of the subjects were collected and genomic DNA was extracted from peripheral blood leucocytes employing the salting-out DNA isolation method. The first exon of the AR gene was amplified by PCR and the primer was fluorescently labeled with FAM. Genotyping of the (CAG) n repeat polymorphisms was performed using capillary electrophoresis and the results were verified by Sanger sequencing method. The exact (CAG) n repeat number was estimated by genotyping results. The genotype frequencies for the AR gene (CAG) n repeat between PCa group and control group was compared. The association between (CAG) n repeat polymorphisms and age, BMI, PSA, Gleason score, tumor stage of PCa patients were also evaluated. Means between PCa group and control group were compared using Student’s t-test or Mann-Whitney U test. Pearson chi-square test was used to compare genotype frequencies between the two groups. The odd ratios (OR) corresponding to95%confidence interval (95%CI) were calculated. A P value<0.05was considered statistically significant. Statistical analysis was carried out using the SPSS19.0computer package.Result A total of323subjects with full data were included in the final analysis,126in the PCa group and197in the control group. There was no significant difference of age and BMI between PCa group and control group (P>0.05). The mean number of AR gene (CAG) n repeats in the PCa group was significantly shorter than in the control group, which were19.81±3.30and20.58±2.75respectively (P<0.05). Compared with control group, the frequence of short (CAG) n repeat (<20) AR gene was significantly higher in the PCa group (OR=1.70,95%CI:1.06,2.71, P<0.05). Short (CAG)n repeat was also a risk factor of high grade (Gleason score>7) tumor (OR=4.12,95%CI:1.84,9.23, P<0.01). However, no significant associations were detected between the AR gene (CAG) n repeat polymorphisms and age, BMI, PSA and tumor stage within PCa group (P>0.05). Conclusions The AR gene (CAG) n repeat polymorphisms were correlated with PCa risk among northern Chinese men. Short (CAG) n repeat (<20) may contribute to PCa susceptibility and high grade tumor. Section2Correlation of androgen receptor gene (CAG)n repeat polymorphisms with the risk of prostate cancer among multi-population:A Meta-analysisObjectives To access the correlation between androgen receptor (AR) gene (CAG) n repeat polymorphisms and prostate cancer (PCa) risk among multi-population using meta-analysis method.Materials and methods Journal articles on the association between AR gene (CAG) n repeats and PCa risk were identified from database published up to August2013. Articles were selected with inclusion/exclusion criteria, and the study of section one was added for meta-analysis. Information from all available studies was extracted blindly and independently by two authors. Stata version12.0was used for the meta-analysis. Q statistic wad performed and subgroup analysis was conducted to expore the source of heterogeneity. Effect size was pooled using Mantel-Haenszel fixed-effect model or random-effects model, and the risk of PCa was assessed by calculating the pooled odd ratio (OR) and the95%confidence interval (95%CI). Ethnicity was stratified for the subgroup analysis. Sensitivity and cumulative analysis were conducted to evaluate the quality and consistency of the results. The potential publication bias was measured using the Begg’s test and Egger’s test and funnel plot were performed. If publication bias existed, the non-parametric’trim and fill’method was used to adjust for it.Results31of215studies and our study were included in this meta-analysis. Caucasians, Asians, Africans and Hispanics were included in the study population. In total, there were5,516PCa patients and6,834healthy controls. Heterogeneity was observed (I2=76.2%, P<0.05) among studies and was greater within Asians (I2=81.5%) and Africans (I2=88.0%). Pooled effects were estimated by using the random-effects model. Short (CAG) n repeat AR gene significantly increase the risk of PCa (OR=1.61,95%CI:1.34,1.94). Subgroup analysis showed that the contribution of shorter (CAG) n repeat on the susceptibility of PCa was predominant in Caucasians (OR=1.37,95%CI:1.15,1.63) and Asians (OR=2.10,95%CI:1.38,3.19). No significant correlations between (CAG) n repeat and PCa risk in Africans and Hispanics were detected. The result of meta-analysis was proved stable and robust through both sensitivity and cumulative analysis. Although significant publication bias was observed by Begg’s and Egger’s test (P<0.05), there was no significant difference for pooled ORs before and after using’trim and fill’method. Conclusions Ethnicity plays an important role on the correlation between the AR gene (CAG) n repeat polymorphisms and PCa risk. Short (CAG) n repeat may increase the PCa risk in Caucasians and Asians but not in Africans and Hispanics.更多还原