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

作　　者：徐蓉[1] 胡刚[2] 林雅军[2] 杨晓姗[1] 柳江 马桂蕾[1] 魏洁[2] 程建华[2] 孙洪范[1] 

机构地区：[1]中国医学科学院北京协和医学院生物医学工程研究所,天津市生物医学材料重点实验室,天津300192 [2]卫生部北京医院/卫生部北京老年医学研究所,卫生部老年医学重点实验室,北京100730 [3]江西省南昌市第三医院内分泌代谢科,330009

出　　处：《国际生物医学工程杂志》2013年第4期193-197,I0002,共6页International Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目（81001439）;中央高校基本科研业务费资助项目

摘　　要：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.

关 键 词：P66SHC 宫颈癌 HELA细胞 增殖 

分 类 号：R737.33[医药卫生—肿瘤] Q813[医药卫生—临床医学]