摘要
目的:观察胰岛素样生长因子-Ⅰ受体(IGF-ⅠR)基因特异的siRNA对胃癌细胞体外增殖及凋亡的影响。方法:设计并体外合成2条靶向IGF-ⅠR的siRNAs,脂质体法瞬时转染MGC803细胞,Western Blot法检测IGF-ⅠR蛋白表达,MTT法检测细胞活力,细胞计数并描记生长曲线,流式细胞仪(FCM)检测凋亡。结果:转染后48h,Western Blot检测显示干扰组(siRNA2-L组、siRNA2-H组、siRNA1组)IGF-ⅠR蛋白抑制率分别为64.41%±4.11%、74.14%±6.15%、89.8%±4.10%;siRNA1组转染后第2～5天细胞活力逐渐减少(分别达49.9%±1.2%、45.9%±4.4%、39.1%±5.1%、29%±4.0%);同期生长曲线显示细胞数分别为对照组的65.58%±4.89%、55.59%±0.82%、44.18%±3.17%、21.15%±1.1%;但FCM检测各组细胞凋亡没有显著差异。结论:特异的siRNAs可显著抑制胃癌细胞中的IGF-ⅠR基因的表达,主要是通过抑制增生而不是增加凋亡导致的。
Objective： To observe the expression of IGF- ⅠR on human gastric cancer MGC803 cells inhibited by special siRNAs and investigate the following effects of the cell proliferation and apoptosis. Methods： Two different siRNAs targeted to IGF- ⅠR were designed ,synthesized and transfected into MGC803 cells, the changes of IGF- ⅠR protein level were detected by Western Blot at 48 h after transfection, then the cell proliferation were examined by MTT, the growth curve was obtained and the apoptosis was detected by Flow Cytometry （FCM）. Results： The 48th hour after transfection, the inhibition ratio of IGF- ⅠR protein of interfering groups （siRNA2-low dose group, siRNA2-high dose group, siRNA1 group） were 64.41%±4.11%,74.14%±6.15%,89.8%±4.10%; On the 2-5 day after transfection, the cell proliferation gradually decreased by 49.9%±1.2%, 45.9%±4.4%, 39.1%±5.1%, 29%±4.0% in siRNA1 group; the cell number decreased by 65.58%±4.89%, 55.59%±0.82%, 44.18%±3.17%, 21.15%±1.1% at the same time; but the early-stage cell apoptosis didn＇t show differences among the groups. Conclusion： Special siRNAs can remarkably inhibit the expression of IGF-ⅠR in gastric cancer cells, the cell survival decreased mainly through the inhibition of the cell proliferation, but not the increasing of the cell apoptosis.
出处
《天津医科大学学报》
2008年第2期157-159,163,共4页
Journal of Tianjin Medical University
基金
天津市科学技术委员会资助项目(05KYZ79)
关键词
小干扰RNA
胰岛素样生长因子-Ⅰ受体
胃癌细胞
Small interfering RNA
Insulin-like growth factor - Ⅰ receptor
Gastric cancer cells
