上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (2): 125-.

• 专题报道（儿科学研究） • 上一篇    下一篇

Fas诱导MML-1凋亡时活化caspase-3 表达与细胞周期的关系

林 芊, 吴伟岚, 卫敏江, 沈 加, 谈 珍, 施 君, 董华腾, 李玉峰   

  1. 上海交通大学 医学院新华医院儿内科, 上海 200092
  • 出版日期:2010-02-25 发布日期:2010-02-25
  • 通讯作者: 李玉峰, 电子信箱: mieuniversity@hotmail.com。
  • 作者简介:林 芊（1978—）, 女, 主治医师, 硕士生；电子信箱: epona913@hotmail.com。
  • 基金资助:

    上海交通大学医学院基金（09XJ076）

Relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of MML-1 cells

LIN Qian, WU Wei-lan, WEI Min-jiang, SHEN Jia, TAN Zhen, SHI Jun, DONG Hua-teng, LI Yu-feng   

  1. Department of Pediatrics, Xinhua Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200092, China
  • Online:2010-02-25 Published:2010-02-25
  • Supported by:

    Shanghai Jiaotong University School of Medicine Foundation, 09XJ076

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摘要：

目的 研究Fas诱导的B淋巴细胞瘤细胞株MML-1凋亡过程中，细胞内活化caspase-3表达与细胞周期的关系。方法 抗Fas抗体孵育MML-1不同时间以诱导细胞凋亡，流式细胞仪检测凋亡率并验证MML-1对Fas诱导凋亡的敏感性。采用PI-Triton X和FITC-active caspase-3双染色分析经Fas诱导凋亡的MML-1内活化caspase-3的表达。以细胞周期蛋白cyclin A、B1、E标记Fas诱导凋亡的处于不同增殖周期MML-1（S、G2/M和G1期细胞），流式细胞术检测细胞内活化caspase-3的表达。结果 Fas诱导6 h后，MML-1凋亡率达到56%。Fas诱导4 h后，活化caspase-3在G1期细胞内表达显著升高，仅有少量S期和G2/M期细胞表达活化caspase-3。Cyclin A、B1阴性而cyclin E阳性MML-1表达活化caspase-3。结论 Fas诱导MML-1凋亡过程中，细胞内活化caspase-3的表达与细胞周期有关。G1后期和S早期的MML-1最先表达活化caspase-3，且对Fas诱导凋亡的敏感性最高。

关键词: 细胞凋亡, caspase-3, 细胞周期, Fas

Abstract:

Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time, and cell apoptosis was induced. Cell apoptotic rates were analysed by flow cytometry, and sensitivity of MML-1 cells to apoptosis was determined. The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3. Cyclin A, B1 and E were selected as cell cycle markers for S, G2/M and G1 phase of MML-1 cells, and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h. After induction by Fas for 4 h, the active form of caspase-3 was mainly expressed in cells of G1 phase, while rarely in cells of S and G2/M phase. Cells with negative cyclin A and B1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent. Cells entering into late G1 and early S phase first express active form of caspase-3, and their sensitivity to Fas-mediated apoptosis is the highest.

Key words: apoptosis, caspase-3, cell cycle, Fas