人p66Shc重组腺病毒抑制Hela细胞增殖的体外研究

Inhibitive effect of human p66Shc recombinant adenovirus on proliferation of Hela cells

目的探讨人p66Shc重组腺病毒抑制人宫颈癌Hela细胞增殖的作用及机制。方法Hela细胞分为空白对照组（不感染腺病毒）、阴性对照组（感染AdenoX—LacZ）和实验组（感染AdenoX—p66Shc）。通过细胞生长曲线、MTT检测分析观察细胞的生长状态；DHE染色法检测细胞内活性氧生成；流式细胞仪检测细胞周期；WesternBlot法检测细胞内相关蛋白的表达。结果p66Shc重组腺病毒能够抑制Hela细胞增殖，且随着p66Shc重组腺病毒处理时间延长和剂量增加其抑制作用增强。与空白和阴性对照组比较，p66Shc重组腺病毒感染48h后，G2／M期细胞比例明显升高，其差异具有统计学意义（P〈0．01）；同时引起Hela细胞活性氧水平显著上升，p53、CyclinB1蛋白表达显著升高，其差异具有统计学意义（P〈0．05）。结论p66Shc重组腺病毒能够显著抑制Hela细胞增殖，并诱导其发生G2／M期阻滞。

Objective To investigate the inhibitive effect and mechanism of human p66Shc recombinant adenovirus on the proliferation of human cervical cancer Hela cells. Methods The cells were divided into three groups： blank control group （Hela cells）, negative control group （Hela cells infected with Adeno X-LacZ） and experiment group （Hela ceils infected with Adeno X-p66Shc）. The proliferation ability was investigated by cell growth curve, MTT detection. Reactive oxygen species （ROS） were detected by Fluorescent Probe-DHE（dihydroethidium）. The cell cycles were analyzed by flow cytometry. Western blot were used to observe associated protein expression. Results The human p66Shc recombinant adenovirus could efficiently suppress the proliferation of Hela cells. The inhibition effects seem to be positive dose and time of action dependent. Compared with blank and negative control group, the ratio of the cell phase G2/M of Hela cells infected with Adeno X-p66Shc for 48h showed a significant increase. The cellular ROS, the expression of p53 and Cyclin B1 proteins were upregulated in Hela cells. Conclusion Adeno X- p66Shc can effectively inhibit proliferation of Hela cells and induce cell cycle arrest at G2/M phase.