小分子RNA干扰同时沉默表皮生长因子受体和胰岛素样生长因子-1受体基因对肝癌细胞周期和凋亡的影响

目的 观察小分子RNA干扰(riRNA)同时沉默表皮生长因子受体(EGFR)和胰岛素样生长因子-1受体(IGF-1R)基因对肝癌细胞周期和凋亡的影响.方法 构建真核表达载体,转染质粒48 h后通过流式细胞仪、噻唑蓝(MTT)检测细胞周期、凋亡、增殖变化以及Western blot检测CDK1、CDK2和p53蛋白的表达.结果 转染48 h后双基因干扰组吸光度为0.2,G0/G1和G2/M期细胞比例分别为72.70±0.26和7.38±0.06,凋亡率为17％,CDK1、CDK2蛋白的表达降低,与对照组和单基因干扰组比较,差异有统计学意义(P＜0.05).结论 同时沉默EGFR和IGF1R基因能有效干扰肝癌细胞增殖、诱导细胞凋亡,并使肝癌细胞阻滞于G0/G1期,干扰多个受体分子可能是一种新的肝癌治疗途经.

Effect of simultaneous silencing of both epidermal growth factors receptors and insulin-like growth factors-1 receptors by small interfering RNA on cycle and apoptosis of hepatocellular carcinoma cells

Objective To explore the effects of simultaneous silencing of both epidermal growth factors receptors (EGFR) and insulin-like growth factors-1 receptors (IGF1R) by small interfering RNA (siRNA) on cycle and apoptosis of hepatocellular carcinoma cells.Methods The expression vectors of IGF1R and EGFR specific for siRNA were constructed,and the recombinant plasmid was stably transfected into human hepatocellular carcinoma (HCC) HepG2 cells with lipofeetion after 48 h.The proliferation of HepG2 cells was determined by methyl thiazol tetrazolium (MTT) assay.Cell cycle distribution was analyzed by using flow cytometry.Western blotting was performed to detect the expression of cyclin-related proteins CDK1,CDK2 and p53.Results The absorbance in siRNA-EGF&IGF1R groups was 0.2 at 48 h after transfection,number of cells in the G1/S and G2/M phase was 72.70 ±0.26 and 7.38 ±0.06 respectively,apoptosis rate was 17％,and the expression of CDK1 and CDK2 was also decreased,which were significantly different from those in normal control,siRNA-HK,siRNA-EGFR and siRNA-IGFI R groups (P ＜ 0.05 ).Conclusion Inhibition of IGF1R and EGFR by siRNA could sharply induce proliferation inhibition and apoptosis,and arrest cell cycle in G0/G1 phase in human HCC cell lines.Conbination of IGF1R and EGFR inhibition may be a promising novel treatment approach for HCC.