摘要
以Molt-4、Jurkat细胞株和外周血淋巴细胞(peripheralbloodlymphocyte,PBL)为靶细胞,检测细胞膜上Fas的表达。人重组Fas配体(recombinanthumanFasligand,rhFasL)诱导细胞6～36h后用改良后的API等方法检测细胞凋亡及诱导凋亡过程中细胞周期蛋白的变化,探讨Fas介导的细胞凋亡与细胞周期的关系。结果显示:rhFasL诱导Molt-4、Jurkat细胞株和植物血凝素刺激进入细胞周期的PBL的凋亡具有细胞周期特异性并始动于G1期;而G0期PBL的细胞膜上虽然也有Fas的表达,但不能诱导细胞凋亡。研究还发现rhFasL诱导细胞凋亡时G1期的细胞周期蛋白D3明显升高,细胞周期蛋白E明显下降。以上结果表明rhFasL体外诱导的细胞凋亡发生在晚G1期,细胞凋亡的发生与细胞是否通过限制点进入细胞周期有关,细胞凋亡发生于晚G1期是G1期细胞周期蛋白E的下降和检测点的监督导致DNA受损的细胞不能通过G1/S交界的结果。
The target cells—leukaemia cell lines （Molt-4 and Jurkat） and peripheral blood lymphocyte were incubated with recombinant human Fas ligand for 6 to 36 hours, apoptosis was detected by sub-G1, traditional annexin-V/PI and modified API methods. Meanwhile, Fas expression in such cells＇ membrane was labelled and the rule of cyclin expression in Fas-mediated apoptosis was studied as well, so as to clarifying the correlation of Fas-mediated apoptosis and cell cycle progression. The data showed that apoptosis induced by recombinant human Fas ligand was cell cycle specific and initiated at G1-phase in the leukaemia cell lines and activated peripheral blood lymphocyte stimulated by phytohemagglutinin, whereas apoptosis in peripheral blood lymphocyte at G0-phase could not be induced effectively. Moreover, cyclin D3 expression increased and cyclin E expression decreased evidently during the induction of apoptosis while the expression of cyclins A and B 1 was unaltered. These findings indicated that Fas-mediated apoptosis was located at late G1-phase and determined by whether the target cells had passed through the restriction point to cell division cycle or not. And the cell cycle specificity of Fas-mediated apoptosis was the result of the cells with DNA damage being blocked at late G1-phase, due to the decrease of cyclin E expression and under the surveillance of G1-phase check point.
出处
《细胞生物学杂志》
CAS
CSCD
2007年第1期109-114,共6页
Chinese Journal of Cell Biology
基金
国家重点基础研究发展规划(973计划)肿瘤项目(No.2004CB518705
No.2002CB513100-2)
国家自然科学基金(No.30440024
No.30570908)
卫生部临床重点项目"临床肿瘤的细胞周期诊断与分析第三期"资助~~
