RNA干扰下调KMT2C基因表达对肝癌细胞HepG2增殖的影响及其机制

Effect of down-regulating histone methyltransferase gene KMT2C with shRNA interference on the proliferation of hepatocellular carcinoma cell line HepG2 and its mechanism

目的探讨组蛋白甲基化转移酶基因KMT2C对肝癌细胞HepG2的增殖影响及其可能的机制。方法采用shRNA基因干扰技术抑制HepG2细胞中KMT2C基因的表达,采用细胞计数、CCK-8细胞增殖实验以及平板克隆形成实验检测细胞增殖能力,流式细胞仪检测细胞周期变化情况;Western blotting和qRT-PCR检测KMT2C基因干扰前后HepG2细胞中EGFR的表达情况。结果甲基化转移酶KMT2C基因干扰成功后,HepG2细胞的增殖能力受到明显抑制(P<0.05),细胞周期阻滞在S期;同时细胞内的EGFR蛋白表达量显著下降(P<0.05)。结论干扰KMT2C基因的表达可以显著抑制肝癌细胞HepG2的增殖,其作用机制可能和EGFR信号通路有关。

Objective To investigate the effect of down-regulating histone methyltransferases gene KMT2C with shRNA interference on the proliferation of hepatocellular carcinoma cell line HepG2 and its mechanism.Methods RNA interference method was used to down-regulate KTM2C expression in HepG2.The cell proliferation was measured by cell counting,CCK-8 kit and colony formation assay and flow cytometry were performed to analyze the cell cycle.Western blotting were used to detect the expression of KMT2C and EGFR in HepG2 before and after the RNA interference.Results After successful down-regulation of KMT2C,the proliferation of HepG2 was obviously inhibited(P<0.05)and the cell cycle was blocked in S phase.At the same time,the EGFR protein expression level was significantly decreased(P<0.05).Conclusion Down-regulation of KMT2C can inhibit the proliferation of HepG2,which the EGFR signal pathway may be involved.