目的 研究肝细胞癌中Wnt/β-连环素信号传导通路与caspase-3、X连锁凋亡抑制蛋白(XIAP)、葡萄糖调节蛋白78(Grp-78)、热休克蛋白(HSP)27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HepG2细胞中沉默β-连环素基因表达,于72、96h提取蛋白质,用Western blot法检测β-连环素、caspase-3、XIAP、Grp-78、HSP27蛋白质的表达.用方差分析的方法进行统计学分析.结果 对照组、转染72h组、转染96h组β-连环素蛋白质表达的灰度值分别为18.9、1.5和3.2 ; β-肌动蛋白表达的灰度值分别为41.1、45.6和47.8 ;caspase-3蛋白质表达的灰度值分别为49.8、5.2和45.1 ; p-caspase-3蛋白质表达的灰度值分别为16.0、67.7和16.3 ; XIAP蛋白质表达的灰度值分别为28.2、21.9和49.9 ; Grp-78蛋白质表达的灰度值分别为23.3、49.9和26.8 ; HSP27蛋白质表达的灰度值分别为29.6、34.8和35.6 ; β-肌动蛋白表达的灰度值分别为32.1、33.6和34.2.针对β-连环素的siRNA转染肝癌HepG2细胞72h和96h均可抑制β-连环素蛋白质的表达(F=160.72,P＜0.01),而96h比72h的表达略有增加,差异有统计学意义.caspase-3的蛋白质表达于72 h被抑制,96h升高恢复至原有水平(F=136.10,P＜0.01);而p-caspase-3的蛋白质表达于72h增加,96h减少至原有水平(F=98.65,P＜0.01).XIAP的蛋白质表达于72 h被抑制,96h表达升高(F=37.29,P＜0.01).Grp-78的蛋白质表达于72 h增加,96h减少至原有水平(F=58.72,P＜0.01).HSP27的蛋白质表达转染前后一致,差异无统计学意义.结论 HepG2细胞中,Wnt/β-连环素信号传导通路与caspase-3、XIAP、Grp-78蛋白质的表达有关,而与HSP27的蛋白质表达无关.Wnt/β-连环素信号传导通路正是通过调节这些因子参与HepG2细胞凋亡及增殖、分化等调节.%Objective To investigate the relationship and significance of Wnt/β-catenin signaling pathway with caspase-3, XIAP, HSP27and Grp-78. Methods The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against β-catenin. After 72 and 96 h, protein was extracted and the protein expressions of β-catenin, caspase-3, XIAP, Grp-78 and HSP27 were detected by Western blot. Results β-catenin protein expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (F = 160.72, P ＜ 0.01). Interestingly, Caspase-3 protein expression was decreased at 72 h and increased to normal at 96 h (F = 136.10, P ＜ 0.01), while p-caspase-3 protein expression increased at 72 h and decreased to normal at 96 h (F= 98.65, P ＜ 0.01). XIAP protein expression decreased at 72 h (F= 37.29, P ＜ 0.01)and increased at 96 h. Grp-78 protein expression increased at 72 h and decreased to normal at 96 h (F = 58.72, P ＜ 0.01). HSP27 protein expression showed no change following transfection (F = 1.91, P ＞ 0.05). Conclusions Wnt/β-catenin signaling pathway is related to the protein expressions of caspase-3, XIAP and Grp-78, but not related to HSP27 protein expression in HCC. Wnt/β-catenin signaling pathway may participate in the regulation of HCC apoptosis, proliferation and differentiation through affecting these factors.
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