SMARCC1基因对肝癌细胞HepG2增殖和凋亡的调控作用

SMARCC1 Gene Inhibits Proliferation and Apoptosis of Hepatocellular Carcinoma Cell Line HepG2

目的探讨下调SMARCC1基因对肝癌细胞Hep G2增殖和凋亡的影响及其可能机制。方法采用siRNA法下调Hep G2细胞的SMARCC1基因,实验分为SMARCC1下调组和对照组,采用MTT法检测两组细胞24 h、48 h、72 h的增殖活性,采用流式细胞术检测两组细胞凋亡,采用RT-PCR法和Western bloting检测两组细胞caspase-3和Bcl-2基因mRNA和蛋白表达水平变化。结果 SMARCC1下调组Hep G2细胞24 h、48 h、72 h增殖活性均低于对照组,SMARCC1下调组晚期凋亡率(7. 3±0. 9%)高于对照组晚期凋亡率(1. 3±0. 1%),P <0. 01。RT-PCR实验结果表明,实验组Caspase-3的mRNA相对表达量高于对照组,实验组Bcl-2mRNA相对表达量低于对照组,均差异有统计学意义。Western blotting实验结果表明,实验组Caspase-3蛋白表达相对灰度值比高于对照组,实验组Bcl-2蛋白表达相对灰度值比低于对照组,均差异有统计学意义。结论下调SMARCC1能抑制肝癌细胞Hep G2的活性,促进其凋亡,其机制与caspase-3和Bcl-2通路有关。

Objective To investigate the effect of down-regulation of SMARCC1 gene on proliferation and apoptosis of human hepatocellular carcinoma cell line Hep G2 and its possible mechanism. Methods SMARCC1 was down-regulated by siRNA method in Hep G2 cells. The experiment was divided into SMARCC1 down-regulated group and the control group. MTT method was used to detect the proliferation activity 24 h,48 h,72 h after the transfection in the 2 groups. Flow cytometry was used to detect the apoptosis by in the 2 groups of cells. RT-PCR method and Western bloting was used to detect the mRNA and protein of Caspase-3 and Bcl-2,respectively. Results The proliferative activity of 24 h,48 h and 72 h in SMARCC1 down-regulation group was lower than that of the control group,and the late apoptosis rate in SMARCC1 down-regulation group( 7. 3 ± 0. 9%) was higher than that of the control group( 1. 3 ± 0. 1%),P < 0. 01. RT-PCR results showed that the relative expression of Caspase-3 in the experimental group was higher than that of the control group,and the relative expression of Bcl-2 mRNA in the experimental group was lower than that of the control group. The results of Western blotting showed that the relative gray value of Caspase-3 protein expression in the experimental group was higher than that of the control group,and the relative gray value ratio of Bcl-2 protein in the experimental group was lower than that of the control group,and the difference was statistically significant. Conclusion Down regulation of SMARCC1 can inhibit the activity of Hep G2 and promote apoptosis,and the mechanism is related to caspase-3 and Bcl-2 pathway.