基础医学与临床 ›› 2010, Vol. 30 ›› Issue (9): 971-975.

• 研究论文 • 上一篇    下一篇

Caveolin-1过表达诱导喉鳞癌HEp2细胞凋亡

顾栋桦 平金良 陈琦 朱荣   

  1. 湖州师范学院附属中心医院 复旦大学上海医学院 复旦大学上海医学院
  • 收稿日期:2009-10-23 修回日期:2009-12-29 出版日期:2010-09-05 发布日期:2010-09-05
  • 通讯作者: 顾栋桦

Overexpression of caveolin-1 induces apoptosis in laryngeal squamous cell carcinoma HEp2 cell line

Dong-hua GU, Jin-liang PING, Qi CHEN, Rong ZHU   

  1. Affiliated Hospital of Huzhou Teachers College Shanghai Medical College, Fudan University
  • Received:2009-10-23 Revised:2009-12-29 Online:2010-09-05 Published:2010-09-05
  • Contact: Dong-hua GU,

摘要: 目的 探讨caveolin-1对喉鳞状细胞癌细胞株HEp2凋亡的影响。方法 通过脂质体法把caveolin-1基因导入喉鳞癌细胞株HEp2细胞,筛选出稳定高表达caveolin-1的克隆,用荧光实时定量反转录-聚合酶链反应(Real-time RT-PCR)和免疫印迹法鉴定;流式细胞仪检测细胞周期与凋亡;Real-time RT-PCR检测survivin的mRNA水平;免疫印迹法检测细胞bcl-2和survivin蛋白的表达;用分光光度法方法检测caspase-3相对活性。结果 成功筛选到稳定高表达caveolin-1的克隆,命名为HEp2-CAV1;与对照组相比,更多的HEp2-CAV1细胞处于G0/G1期,凋亡率增加,caspase-3相对活性水平上升2.72倍(p<0.05),survivin mRNA的表达水平减少74%(p<0.01),survivin与Bcl-2蛋白表达水平明显减少。结论 caveolin-1能够促进喉鳞状细胞癌细胞株HEp2的凋亡, bcl-2和survivin的表达下调可能是其促凋亡的重要机制之一。

关键词: 喉肿瘤, 质膜微囊蛋白, 凋亡

Abstract: Objective To investigate the effects of caveolin-1 on apoptosis of laryngeal squamous cell carcinoma cell line HEp2. Methods Human caveolin-1 gene were transfected into HEp2 cell line, the positive clones with high expression of caveolin-1 were identified by fluorescence quantitative real time reverse transcriptase-polymerse chain reaction(Real-time RT-PCR)and Western Blot. Cell cycle and apoptosis were analyzed by Flow cytometry. Caspase-3 relative activities were calculated according to the changs of optical density after reaction with caspase-3 substrate. Real-time RT-PCR was used to determine the expression level of survivin mRNA and the expression of bcl-2 and survivin protein were detected by Western Blot. Results the clones stably overexpressing caveolin-1 were obtained, namely HEp2-CAV1. Overexpressing caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptosis cell fraction. Compared with the parental HEp2 cells, the realtive activites of caspase-3 increased by 2.72 fold(p<0.05), the expression level of survivin mRNA reduced by 74%(p<0.01) and the protein bcl-2 significantly decreased in HEp2-CAV1 cells, respectively. Conclusion overexpression of caveolin-1 induces apoptosis in laryngeal squamous cell carcinoma HEp2 cell line, downregulation of bcl-2 and survivin expression may be involved in its mechanism.

Key words: laryngeal tumor, caveolin, apoptosis