IGF-1R表达下调抑制肾癌786-0细胞的增殖、迁移、侵袭和细胞周期G1/S期转变

摘要

目的:观察小干扰RNA (siRNA)介导的胰岛素样生长因子1受体(IGF-1R)基因沉默对肾癌细胞786-0增殖、迁移、侵袭和细胞周期的影响.方法:采用实时定量PCR(real-time PCR)和蛋白印迹法(West-ern blot)测定转染后肾癌细胞IGF-lR及下游相关产物的表达;采用Cell counting kit-8(CCK-8)检测细胞增殖能力;Transwell实验及划痕实验检测细胞迁移及侵袭能力;流式细胞仪检测细胞凋亡及周期改变.结果:IGF-1R基因的siRNA可有效抑制IGF-1R mRNA及蛋白的表达(P＜0.01);细胞增殖实验结果显示IGF-1R-siRNA转染组增殖能力显著减弱(P＜0.01);Transwell实验及划痕实验结果显示IGF-1R-siRNA转染组侵袭及迁移能力明显降低(P＜0.01);流式细胞术检测到转染后细胞G1期细胞数明显升高,发生G1/S期阻滞(P＜0.05);IGF-1R下游蛋白p-IRS1、p-IRS2、p-SHC水平显著降低.结论:IGF-1R表达下调可以抑制肾癌786-0细胞的生长、增殖、迁移和侵袭的能力,并可以使肾癌786-0细胞G1/S期细胞周期停滞.%Objective:To explore the effects of small interference RNA (siRNA)-mediated knockdown of IGF-1R on cell growth,proliferation,invasion,migration and cell cycle phase G1/S transition in 786-0 RCC cells.Methods:Real-time PCR and Western blot were used to detect the expression of IGF-1R and the downstream proteins of IGF -1R pathway.Cell proliferation ability was determined by CCK-8 method,the change of cell cycle was detected by flow cytometry.Results:RNA interference targeting insulin-like growth factor receptor 1 (IGF-1 R) could effectively inhibit the expression of mRNA and protein of IGF-1R (P ＜ 0.01).Cell proliferation assay showed that the proliferation of transfected 786-0 cells were significantly decreased (P ＜ 0.01).Transwell assay and Wound-healing assay revealed that the abilities of migration and invasion were significantly decreased in 786-0 cells transfected with IGF-1R-siRNA (P ＜ 0.01).The transfected 786-0 cells had a significant increase in G1-phase population (P ＜0.05).Western blotting showed that 786-0 cells transfected with IGF-1R-siRNA decreased the expression of p-IRS1,p-IRS2 and p-SHC.Conclusion:The down-regulation of IGF-1R suppressed the proliferation,migTation and invasion and induces G1/S phase cell cycle arrest in 786-0 RCC cells.