基因多态性的交互作用与食管癌患者血浆凝血酶活性及病理分期的关系

摘要

目的 探讨凝血酶原基因3'非翻译区G20210A与组织因子途径抑制物(TFPI)基因5'非翻译区C399T多态性的交互作用与食管癌患者血浆凝血酶活性及病理分期的关系.方法 依据TNM分期,选择新乡医学院第一附属医院2011年5月至2015年8月收治的食管癌TNM Ⅰ期～Ⅳ期及TNM 0期患者各198例,发色底物法测定血浆凝血酶活性,并应用PCR RFLP技术检测各组患者的外周血白细胞的凝血酶原基因3 '非翻译区G20210A和TFPI基因5'非翻译区C399T多态性,采用非条件Logistic回归分析估算凝血酶原基因3'非翻译区G20210A和TFPI基因5'非翻译区C399T与食管癌浸润与转移风险的调整比值比(OR)及95％可信区间(95％CI),并分析二者多态性的交互作用对血浆凝血酶活性和食管癌浸润与转移的影响.结果 Ⅰ期组G20210A(GA)、G20210A(AA)、C399T (CT)和C399T(TT)基因型频率分别为24.24％、26.77％、24.24％和25.76％,Ⅱ期组分别为34.34％、37.37％、34.85％和36.36％,Ⅲ期组分别为39.90％、42.93％、40.41％和41.92％,Ⅳ期组分别为45.45％、46.97％、45.35％和46.46,对照组分别为13.64％、14.14％、13.13％和13.64％,各分期组与0期对照组之间差异均有统计学意义(P均＜0.01).G20210A(GA)和G20210A(AA)基因型患者的食管癌浸润与转移的风险均显著增加,C399T(CT)和C399T(TT)基因型者食管癌浸润的风险也显著增加.基因突变的协同分析发现,G20210A(AA)/C399T(TT)基因型者各分期组和0期组的分布频率分别为7.07％、14.14％、18.18％、21.71％和1.52％,各组之间差异均有统计学意义(P均＜0.01).G20210A(AA)/C399T(TT)基因型者食管癌浸润与转移的风险显著增加,且两基因型在食管癌浸润与转移中存在正向的交互作用(γ均＞1),此外,G20210A(GA)和C399T(TT)之间、G20210A(GA)和C399T(CT)之间及G20210A(AA)和C399T(CT)之间均存在正向交互作用(γ均＞1).各分期患者血浆凝血酶活性明显高于0期组,且各分期组患者之间也有明显差异(P＜0.01).突变基因型携带者的血浆凝血酶活性明显高于在同一TNM分期野生型携带者的血浆凝血酶活性(P＜0.01).结论 G20210A和C399T的基因突变均是食管癌浸润与转移的易患因素,基因突变的交互作用增加了食管癌浸润与转移的风险,这可能与其提高血浆凝血酶活性密切相关.%Objective To investigate the correlation of interaction between polymorphisms of prothrombin gene G20210A in 3' untranslated region and tissue factor pathway inhibitor (TFPI) gene C399T in 5' untranslated region with thrombin activity in plasma and the pathological stages of esophageal carcinoma.Methods Based on TNM method,we selected 198 patients with stage Ⅰ esophageal carcinoma,198 with stage Ⅱ,198 with stage Ⅲ,and 198 with stage Ⅳ from the First Affiliated Hospital of Xinxiang Medical College from May 2011 to August 2015 for this study;198 patients with esophageal carcinoma of stage 0 served as the control group.The thrombin activity in plasma were determined by chromogenic substrate assay.The genetic polymorphisms of prothrombin gene G20210A in 3' untranslated region and TFPI gene C399T in 5' untranslated region in peripheral blood leukocytes of the above-mentioned patients were analyzed by PCR-RFLP technique.Unconditional logistic regression model and single factor analysis were performed to calculate the adjusted odds ratios (OR) and 95％ confidence intervals (95％ CI) of polymorphisms prothrombin gene G20210A and TFPI gene C399T polymorphisms and to analyze the interaction of nucleotide polymorphisms with thrombin activity in plasma and the pathological stages of esophageal carcinoma.Results The frequencies of G20210A (GA),G20210A (AA),C399T (CT) and C399T (TT) were 24.24％,26.77％,24.24％ and 25.76％ in stage Ⅰ group;34.34％,37.37％,34.85％ and 36.36％ in stage Ⅱ group;39.90％,42.93％,40.41％ and 41.92％ in stage Ⅲ group;45.45％,46.97％,45.35％ and 46.46 in stage Ⅳ group;and 13.64％,14.14％,13.13％ and 13.64％ in stage 0 group,respectively.Statistical tests showed significant difference in the frequencies among each group (all P＜0.01).The risks of invasion and metastasis of esophageal carcinoma significantly increased in the subjects with G20210A,in those with G20210A(AA) genotype,in those with C399T (CT) genotype and in those with C399T (TT) genotype.Combined analysis of the polymorphisms showed that percentage of G20210A (AA)/C399T (TT) in stage Ⅰ group,stage Ⅱ group,stage Ⅲ group,stage Ⅳ group and stage 0 group was 7.07％,14.14％,18.18％,21.71％ and 1.52％,respectively,and statistical tests showed significant difference in the frequency among each group (all P＜0.01).People who carried G20210A(AA)/C399T(TT) had higher risks of invasion and metastasis of esophageal carcinoma,and statistical analysis suggested a positive interaction between G20210A (AA) and C399T (TT) in increasing the risks of invasion and metastasis of esophageal carcinoma (All γ＞ 1).Likewise,there were also positive interactions in the pathogenesis of invasion and metastasis of esophageal carcinoma between G20210A (GA) and C399T (TT),G20210A (GA) and C399T(CT),G20210A (AA) and C399T (CT) (All γ＞1).The thrombin activities in plasma in stage Ⅰ,Ⅱ,Ⅲ and Ⅳ groups were all significantly higher than those in stage 0 group,and there were significant differences among stage Ⅰ,stage Ⅱ,stage Ⅲ and stage Ⅳ in thrombin activities (all P＜0.01).Patients with mutation genotype had significantly higher thrombin activities than those with wild homozygous in the same TNM stage.Conclusion G20210A and C399T gene mutations are the risk factors in the invasion and metastasis of esophageal carcinoma.Significant interactions between G20210A and C399T mutations increase the risk of invasion and metastasis of esophageal carcinoma,which may be closely related to their increased thrombin activities in plasma.