Caveolin-1过表达抑制宫颈鳞癌Hela细胞的体外生长研究  

Overexpression of caveolin-1 inhibits the growth of human cervical squamous cell Hela cell line in vitro

作　　者：郑庆玲[1] 顾栋桦[2] 平金良[2] 朱荣[3] 陈琦[3] 

机构地区：[1]湖州师范学院医学院门诊部,浙江省湖州313000 [2]浙江省湖州师范学院附属中心医院病理科 [3]复旦大学上海医学院病理学系

出　　处：《中国医师杂志》2010年第10期1304-1308,共5页Journal of Chinese Physician

基　　金：湖州市科技局科技攻关项目（2008GS07）

摘　　要：目的 探讨转染caveolin-1基因对宫颈鳞状细胞癌Hela细胞株体外生长的影响.方法 用脂质体法把caveolin-1真核表达质粒导入Hela细胞内,筛选出稳定高表达caveolin-1的细胞克隆,并用RT-PCR、细胞免疫荧光和免疫印迹法鉴定.MMT法检测细胞的生长能力,流式细胞仪检测细胞周期分布和凋亡情况,免疫印迹法检测细胞外信号调节激酶（extracellular regulated protein kinases,Erk）1/2的磷酸化水平.结果 转染caveolin-1基因后,成功筛选得到稳定高表达caveolin-1的Hela细胞克隆,与未转染对照组相比,转染后的Hela细胞生长速度明显减慢（P〈0.05）,更多的细胞阻滞在G0/G1期[（68.04±2.57）%vs（53.41±1.01）%],凋亡细胞比例增高[（19.18±2.20）%vs（5.63±0.55）%,P〈0.05],Erk1/2相对磷酸化水平明显下降（0.28±0.05 vs 0.81±0.07,P〈0.05）.结论 caveolin-1基因能够抑制宫颈鳞癌Hela细胞株的生长、促进细胞凋亡;Erk1/2磷酸化水平下降可能在其抑癌机制中发挥重要作用.Objective To investigate the effects of caveolin-1 overexpressing on the growth of cervical squamous cell cancer Hela cell line. Methods Eukaryotic expression vector of human caveolin-1 gene was introduced into Hela cells by Lipofectamine. The clones stably overexpressed caveolin-1 were identiffed by RT-PCR, immunofluorescence cell staining techniques and Westernblotting. Cells proliferation viabihty was tested by MTT assay, and flow cytometry was used to assay the cell cycle and apoptosis, and the relative phosphorylation level of extracellular regulated protein kinases （Erk1/2） were detected by Westernblotting. Results The clones stably overexpressed caveolin-1 were obtained. Compared with the parental Hela cells, the tranfected cells exhibited a slower rate of growth. FAGS analysis results revealed that overexpression of caveolin-1 resulted in the cell cycle arrest in the G0/G1 [ （ 68. 04 ± 2. 57 ） % vs （ 53.41 ±1.01）%] phase and increased the apoptotic cell fraction[ （19. 18 ±2.20）% vs （5.63 ±0.55）%, P 〈0. 05 ]. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of Erk1/2（0.28 ±0.05 vs 0.81 ±0.07, P 〈0.05）. Conclusions Overexpression of caveolin-1 suppressed the growth of Hela cells and induced apoptosis, down-regulation of Erk1/2 phosphorylation might be involved in its mechanism.

关 键 词：窖蛋白1/代谢 宫颈肿瘤/代谢 细胞生长过程 

分 类 号：R737.33[医药卫生—肿瘤]