Wnt/β-连环素信号通路对肝细胞癌信号分子的调节作用及其意义

目的 研究肝细胞癌中Wnt/β-连环素信号传导通路与糖原合成激酶(GSK)-3 β、STAT3、Smad3和人端粒酶逆转录酶(TERT)的关系及其意义.方法 用RNAi技术将针对β-连环素的siRNA转染入肝癌细胞系HepG2细胞中沉默β-连环素基因,于72h和96h提取蛋白质,用Western blot法检测β-连环素、GSK-3 β、p-GSK 3 β、STAT3、Smad3和TERT蛋白质的表达.用Student's t检验及方差分析进行统计学分析.结果 针对β-连环素的siRNA转染HepG2细胞72 h和96 h均可抑制β-连环素蛋白质的表达,且96 h比72 h的表达略有增加(t=4.43,P＜0.05),而GSK-3 β及p-GSK-3 β的蛋白质表达于转染后72 h和96 h依次增加(tGSK 3β=4.98,tp-GSK-3β=29.83,P值均＜0.05);STAT3的蛋白质表达于转染前后一致,差异无统计学意义(F=0.49,P＞0.05);smad3的蛋白质表达于转染后72 h增加(t=10.67,P＜0.05),96 h减少至原有水平(与转染前比较,t=0.90,P＞0.05);TERT的蛋白质表达于转染后72 h减少(t=4.18,P＜0.05),96 h增加至原有水平(t=1.26,P＞0.05).结论 肝细胞癌中Wnt/β-连环素信号通路可能通过调节GSK-3 β、p-GSK-3 β、Smad3、TERT蛋白质的表达来参与肝癌的发生和发展过程;而与STAT3蛋白质的表达无关.

The role and significance of Wnt/β-catenin signaling pathway regulating the signaling molecules in hepatocellular carcinoma

Objective To investigate the role and significance ofWnt/β -catenin signaling pathway regulating GSK-3 β, STAT3, Smad3 and TERT in hepatocellular carcinoma (HCC). Methods The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against β -catenin. Proteins were extracted and the expressions of β -catenin, GSK-3 β, p-GSK-3 β, STAT3, Smad3 and TERT were detected by Western blot at 72 h and 96 h respectively after transfection. Results β -catenin expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (t = 4.43, P ＜ 0.05). Interestingly, GSK-3 β and p-GSK-3 β expressions increased gradually at 72 and 96 h (tGSK-3 β = 4.98, tp-GSK-3 β = 29.83, P ＜ 0.05)respectively, and STAT3 expression showed no alteration after transfection (F = 0.49, P ＞ 0.05). Smad3expression was increased at 72 h (t = 10.67, P ＜ 0.05) and decreased to normal at 96 h (t = 1.26,P ＜ 0.05), while TERT expression decreased at 72 h (t = 4.18, P ＜ 0.05) and increased to normal at 96 h (t = 1.26, P ＞ 0.05). Conclusions Wnt/ β -catenin signaling pathway is related to the expressions of GSK-3 β, Smad3 and TERT, but perhaps not related to STAT3 protein expression in HCC. It suggested that Wnt/ β -catenin signaling pathway might participate in HCC genesis and development through regulating the above three factors.