抑制ERBB2基因对肝癌HepG2细胞生长及PI3K/AKT信号通路的影响

Inhibition of ERBB2 gene on hepatoma HepG2 cell growth and PI3K/AKT signaling pathway

目的:探讨抑制ERBB2基因表达对肝癌HepG2细胞生长及PI3K/AKT信号通路的影响。方法:采用脂质体法将ERBB2 siRNA转染至肝癌HepG2细胞,记为实验组(ERBB2-siRNA组),以siRNA对照转染的细胞记为阴性对照组(Con-siRNA组),将不经任何处理的细胞记为空白对照组。采用qRT-PCR和Western blot法检测转染效率,CCK-8法检测细胞增殖能力,克隆形成实验检测细胞克隆形成能力,流式细胞仪检测细胞周期,Western blot法检测HepG2细胞中增殖细胞核抗原(PCNA)、AKT和p-AKT蛋白表达水平。结果:在HepG2细胞中转染ERBB2siRNA能够抑制ERBB2的表达(P<0.05)。与Con-siRNA组比较,抑制ERBB2的表达后细胞生长和克隆形成能力明显被抑制,细胞周期发生G0/G1期阻滞,细胞中PCNA和p-AKT蛋白表达下调,差异均有统计学意义(P<0.05)。结论:抑制ERBB2基因表达可抑制肝癌HepG2细胞的生长,其作用机制可能与PI3K/AKT信号通路的激活有关。

Aim:To investigate the effect of inhibiting the expression of ERBB2 gene on the growth of HepG2 cells and PI3K/AKT signaling pathway.Methods:The ERBB2 siRNA was transfected into HepG2 cells by liposome method,which was recorded as the experimental group(ERBB2-siRNA group).The cells transfected with siRNA control were recorded as negative control group(Con-siRNA group),the cells without any treatment were recorded as blank control group.The transfection efficiency was detected by qRT-PCR and Western blot,the cell proliferation ability was detected by CCK-8 method,the cell clone formation ability was detected by colony formation assay,and the cell cycle was detected by flow cytometry.Proliferating cell nuclear antigen(PCNA),AKT and p-AKT protein levels were measured in HepG2 cells by Western blot.Results:Transfection of ERBB2 siRNA in HepG2 cells inhibited the expression of ERBB2(P<0.05).Compared with the Con-siRNA group,the cell growth and colony formation abilities were significantly inhibited after reducing the expression of ERBB2(P<0.05).The cells arrested at G0/G1 phase and the expressions of PCNA and p-AKT proteins in the cells were down-regulated(P<0.05).Conclusion:Inhibition of ERBB2 gene expression could inhibit the growth of HepG2 cells,and its mechanism is related to the activation of PI3K/AKT signaling pathway.