Fas诱导MML-1凋亡时活化caspase-3表达与细胞周期的关系  

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

作　　者：林芊[1] 吴伟岚[1] 卫敏江[1] 沈加[1] 谈珍[1] 施君[1] 董华腾[1] 李玉峰[1] 

机构地区：[1]上海交通大学医学院新华医院儿内科,上海200092

出　　处：《上海交通大学学报（医学版）》2010年第2期125-128,139,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海交通大学医学院基金(09XJ076)~~

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-TritonX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、B_1、E标记Fas诱导凋亡的处于不同增殖周期MML-1(S、G_2/M和G_1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G_1期细胞内表达显著升高,仅有少量S期和G_2/M期细胞表达活化caspase-3。Cyclin A、B_1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。G_1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。Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fasmediated 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.

关 键 词：细胞凋亡 CASPASE-3 细胞周期 FAS 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]