腺病毒介导p53基因对胰腺癌细胞抑制作用的体外实验研究

The experimental study on inhibitory effect on pancreatic tumor cells by adenovirus - mediated p53 gene

目的探讨腺病毒介导p53基因（adenovims—mediated p53 gene，Ad-p53）对人胰腺癌细胞株PANC-1的生长抑制作用。方法以Western Blot方法检测加入Ad—p53后PANC-1细胞在48h、72hp53表达情况以及未处理组细胞48h、72hp53的表达。采用四甲基偶氮唑盐（MTT）比色实验检测不同MOI值的Ad—p53感染后，PANC-1细胞的生长情况并绘制生长曲线，用PI、annexin V—FITC双重染色并用流式细胞技术检测受感染细胞的凋亡比例。结果Westem Blot结果显示，处理组细胞内48h、72hp53蛋白表达明显高于相同时间点对照组细胞；MTT结果显示不同MOI值细胞生长抑制情况不同，MOI值越高，对细胞生长的抑制程度也越高；流式检测细胞在MOI值为150时，0h、48h、72h凋亡比例分别为（7．5±0．8）％、（22．5±2．3）％和（34．4±2．7）％，对照组细胞为（5．8±0．4）％、（8．3±1．7）％和（9．7±2．1）％。结论Ad-p53能有效感染胰腺癌细胞，导致胰腺癌细胞凋亡。

Objective To investigate the inhibitory effect of adenovirus - mediated p53 gene （ Ad - p53 ） on growth of pancreatic tumor cell line PANC - 1 cells. Methods The expression of p53 protein in Ad - p53 transfected and non - transfected PANC - 1 cells was detected at the 48th, and 72nd h. The MTT method was used to observe the inhibitory effects of Ad - p53 transfection on the growth of pancreatic tumor cells, and a growth curve was drawn. The apoptosis rate of Ad - p53 transfected cells double stained with PI and annexin V - FITC was measured by using flow cytometry. Results The results of Wetern blot showed that the expression of p53 protein in the transfected cells was significantly increased as compared with that in the non - transfected cells at the same time point. At different MOI, the inhibitory rate was different. The higher the MOI was, the higher the inhibitory rate was. The flow eytometry revealed when MOI was 150,the apoptosis rate at 0,48,72 h in Ad -p53 -transfected group was （7.5 ± 0.8 ） %, （ 22.5 ± 2.3 ） %, and （ 34.4 ± 2.7 ） %, and that in non - transfected group was （ 5.8 ± 0.4 ） %, （ 8.3 ± 1. 7） % , and （9.7 ± 2.1 ） % respectively. Conclusion Ad - p53 can effectively infect pancreatic tumor cells and cause apoptosis of PANC - 1 cells.