CAJ | 학술논문

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.
목적 관찰소분자RNA간우(riRNA)동시침묵표피생장인자수체(EGFR)화이도소양생장인자-1수체(IGF-1R)기인대간암세포주기화조망적영향.방법 구건진핵표체재체,전염질립48 h후통과류식세포의、새서람(MTT)검측세포주기、조망、증식변화이급Western blot검측CDK1、CDK2화p53단백적표체.결과 전염48 h후쌍기인간우조흡광도위0.2,G0/G1화G2/M기세포비례분별위72.70±0.26화7.38±0.06,조망솔위17％,CDK1、CDK2단백적표체강저,여대조조화단기인간우조비교,차이유통계학의의(P＜0.05).결론 동시침묵EGFR화IGF1R기인능유효간우간암세포증식、유도세포조망,병사간암세포조체우G0/G1기,간우다개수체분자가능시일충신적간암치료도경.
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.