p27kip1的过表达对肾癌细胞增殖及凋亡的影响

目的:研究外源性 p27kip1 基因对肾癌细胞周期、细胞凋亡及增殖的影响。方法:将含p27cDNA的质粒在脂质体介导下在体外转染肾癌GRC 1细胞,Western Blot、FCM及MTT检测分析外源性 p27kip1 蛋白在细胞内的表达,观察其对细胞周期、凋亡及细胞增殖的影响。结果:体外转染 GRC 1 细胞 48 h 后, p27 蛋白在p27kip1转染后的 GRC 1 细胞中高表达。FCM检测表明,细胞停滞于 G1 期,实验组 G1 期细胞占66 9%,并出现亚G1 凋亡峰;而空白对照组和转染空载体组 G1 期细胞分别为 32 2% 和33 8%。MTT 法检测表明,转染 p27cDNA 后,实验组细胞增殖明显受抑。结论: p27kip1 基因在体外转染GRC 1细胞并过表达p27kip1蛋白,抑制细胞由G1 期向 S期过渡,从而抑制细胞增殖,诱导细胞凋亡。

Effect of overexpression of p27kip1 on renal carcinoma cell proliferation and cell apoptosis

OBJECTIVE:To investigate the effects of exogenous p27kip1 gene on cell cycle and apoptosis. METHODS: The p27kip1 cDNA was transfected into renal carcinoma cell line of human with lipofectamine. After infecting GRC-1 cells for 48 h, the exogenous p27kip1 protein express in GRC-1 cells and its effects on cell cycles and proliferation were determined by Western blot, FCM and MTT assay. RESULTS: Over-expression of p27kip1 protein was observed in p27kip1 cDNA-infectsd cells. As a result,the proliferation of the cells was greatly inhibited, the cells being arrested in G 1 phase and infection of GRC-1 cells resulted in a marked increase in the sub-G 1. CONCLUSIONS: p27kip1 gene can be efficiently transfered into GRC-1 cells and over-express p27kip1 protein in the infected cells. Overexpression of p27kip1 induces G 1 arrest and apoptosis, suppressive effect on cell growth.