腺病毒介导的人突变p27基因转染对人大肠癌细胞系的作用

The Effect of Adenovirus-mediated Human Mutant p27 Gene Transfection on the Proliferation of Human Colorectal Carcinoma Cell Line

目的 用人重组p2 7mt腺病毒感染大肠癌细胞株SW 480 ,研究p2 7mt对大肠癌细胞的抑制作用。方法 用重组腺病毒Ad -p2 7mt转染SW 480后 ,用Westernblot检测目的基因的表达 ;用流式细胞仪对转染细胞进行倍体分析 ,绘制生长曲线观察对细胞的生长抑制作用。结果 Ad -p2 7mt转染SW 480后 ,通过Westernblot分析发现p2 7在细胞中出现了高表达 ;PI染色流式细胞仪检测 ,77 9%的细胞阻滞于G0 /G1 期 ,而Ad -LacZ组及空白对照组分别为 2 5 5 7%和 2 5 2 9%(P <0 0 0 1) ;由生长曲线可以看出Ad -p2 7mt于 2 4h内对细胞就有明显的抑制作用 ( P<0 0 5 ) ,且持续时间长。结论 重组腺病毒能够有效转染p2 7mt基因进入SW480细胞内并获得了高表达 ;p2 7mt对细胞周期有明显的阻滞作用且对细胞的生长有明显的抑制作用。突变p2 7是一种细胞周期高效阻滞因子和高效生长抑制因子 ,为p2

Objective To explore the inhibition effect of recombinant mutant type(mt) p27 gene adenovirus (Ad-mt p27) transfection on the proliferation of colorectal cancer cell line SW480. Methods The recombinant mt p27 adenovirus vector was transfected into SW480 cells, western blotting analysis was applied to determine the p27 expression, and cell cycle distribution was analyzed using flow cytometry, and growth curve was drawn to observe the cell growth. Results There was high expression of p27 protein in the transfected SW480 cells. 77 9% cells were blocked in G 1/G 0 phase in the transfected group, which the percentage of the G 1/G 0 phase of Ad-LacZ transfected group and normal control group were 27 5% and 25 29% respectively. Ad-mt p27 transfection markedly inhibited SW480 cell growth in 24h and the inhibition effects was maintained for 7 days. Conclusion Recombinant adenovirus could efficaciously trasfect mt p27 gene into SW480 cells, and obtain high level mt p27 exprerssion. mt p27 markedly bolcked SW480 cell cycle progression and prominently inhibited SW480 cell growth. The mutant p27 gene transfection may be a new way of gene therapy for colorectal cancer.