摘要目的 探讨PNPLA3、TM6SF2基因多态性及其与吸烟、饮酒交互作用对HBV相关肝癌(hepatitis B virus-associated hepatocellular carcinoma,HBV-HCC)的影响.方法 收集2010年1月至2014年3月HBV-HCC患者、慢性乙型肝炎(乙肝)患者(CHB)、肝硬化患者(LC)以及健康体检者的血液标本,应用飞行质谱技术检测patatin样磷脂酶域3(PNPLA3)基因位点rs738409和6号跨膜超家族成员2(TM6SF2)基因位点rs58542926的单核苷酸多态性(SNP).利用在线SNP stats寻找基因多态性的最优赋值方法;检验SNP是否符合哈-温(H-W)遗传平衡定律;采用多分类logistic回归分析PNPLA3和TM6SF2多态性及吸烟、饮酒因素对HBV-HCC的影响,采用叉生分析和二分类logistic回归分析探讨基因-基因、基因-吸烟、饮酒交互作用对HBV-HCC的影响.结果 H-W遗传平衡检验结果显示,CHB组rs738409位点的基因型频率分布不符合H-W遗传平衡定律(x2=11.980,P＜0.005),CHB组rs58542926位点、HBV-HCC组和LC组rs738409和rs58542926位点均符合H-W遗传平衡定律;调整年龄、性别的影响后,与健康体检者相比,HBV-HCC组rs58542926突变的OR=1.659,95％CI:1.026 ～ 2.684,P=0.039.与CHB组相比,HBV-HCC组饮酒的OR=1.680,95％CI:1.121 ～ 2.519,P=0.012.与LC组相比,HBV-HCC组饮酒与吸烟的OR值(95％CI)分别为1.539(1.071 ～ 2.213)和1.453(1.005 ～ 2.099).交互作用分析显示,HBV-HCC组与CHB+ LC组相比,rs738409与rs58542926交互作用的叉生分析相加模型OR=1.548(U=1.885,P=0.029),logistic回归相乘模型OR=1.658(P=0.024);饮酒与rs738409的交互作用叉生分析,饮酒且rs738409突变相加模型OR=1.811 (U=1.965,P=0.024),相乘模型无统计学意义;logistic回归相乘模型吸烟与饮酒的交互作用OR=1.756 (P＜0.001).结论 TM6SF2基因突变、吸烟、饮酒是HBV-HCC的危险因素,PNPLA3与TM6SF2基因都突变、饮酒并且吸烟是HBV-HCC的危险因素.PNPLA3的单基因突变可以减弱饮酒对HBV-HCC的危害.

abstractsObjective To explore the SNP effects ofpatatin-like phospholipase domain which containing 3 (PNPLA3),transmembrane 6 superfamily member 2 (TM6SF2) gene,environmental effects of smoking,alcohol drinking and interaction between gene-gene,gene-environment and drinking-smoking on hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC).Methods We collected anticoagulant peripheral blood from patients of HBV-HCC,chronic hepatitis B (CHB),liver cirrhosis (LC) and from healthy controls to detect the single nucleotide polymorphism (SNP) of patatin-like phospholipase domain containing 3 (PNPLA3) gene loci rs738409 and transmembrane 6 superfamily member 2 (TM6SF2) gene loci rs58542926,using the flight mass spectrometry method.The optimal assignment value of gene polymorphisms was defined by using the online SNP stats.Hardy-Weinberg (H-W) balance was tested for SNP.Effects of the genetic and environmental factors to HBV-HCC were analyzed by using the multiple classification logistic regression method.The gene-gene,gene-smoking and alcohol drinking interaction effects were investigated by Fork-Life analysis and binary logistic regression methods.Results The frequency distribution of CHB group rs738409 loci seemed not in conformity with the H-W balance (x2=11.980,P＜0.005).Two loci frequency distributions in the other groups were all in accordandce with the H-W balance.After adjusting for influences on age and sex and comparing to the healthy group,the rs58542926 mutation appeared as OR=1.659,95％CI:1.026-2.684,P=0.039,in the HBV-HCC group.When comparing to CHB group,the HBV-HCC group presented that drinking as OR=1.680,95％CI:1.121-2.519,P=0.012.When comparing to the LC group,the ORs of drinking and smoking were 1.539 (1.071-2.213) and 1.453 (1.005-2.099) respectively,in the HBV-HCC group.When comparing to the CHB + LC group,interactions between the HBV-HCC group were found rs738409 and rs58542926 on additive model OR=1.548 (U=1.885,P=0.029) and OR=1.658 (P=0.024) on logistic regression model while drinking was rs738409 on interaction additive model with OR=1.811(U=1.965,P=0.024).As for drinking and mutation of rs738409,the multiplication model of logistic regression showed no statistically significant differences.Interaction between smoking and drinking appeared as OR=1.756 (P＜0.001) in the logistics regression multiplication model.Conclusions Factors as mutation of TM6SF2,smoking and drinking all appeared as risk factors for HBV-HCC.Mutations of both PNPLA3 and TM6SF2,together with smoking and drinking all served as risk factors for HBV-HCC.However,the mutation of single PNPLA3 appeared as a protective factor on HBV-HCC.