体外Fas介导细胞凋亡的细胞周期特异性的检测方法

• 作 者: 何小军 胡静 李小兰 Hui Xiao Deding Tao
• 机 构: 华中科技大学同济医学院附属同济医院;Tongji Cancer Research Institute/Molecular Medical Center, Tongji Hopital, Tongji
• 出 处: 中德临床肿瘤学杂志
• 发表日期: 2006年5期
• 关键词: Fas-mediated apoptosis cell cycle flow cytometry leukaemia cell peripheral blood

Objective: To establish a system in detecting the cell cycle specificity induced by recombinant human Fas ligand in vitro,so as to provide a reliable platform for further exploring the mechanism of cell cycle control and regulation in Fas-mediated apoptosis.Methods: The target cells-leukaemia cell lines and activated peripheral blood lymphocytes stimulated by phytohemagglutinin were incubated with recombinant human Fas ligand for 6 to 36 h,apoptosis was detected by sub-G1,common annexin-V/PI and modified annexin V and propidium iodide (API) methods and analysed by flow cytometry.Results: The modified API method demonstrated that Fas-mediated apoptosis was cell cycle specific and initiated at G1 phase.The common annexinV/PI method showed the most appropriate condition for the detection of typical cell cycle-specific apoptosis.The sub-G1 method could only illuminate late apoptosis and DNA histogram.Conclusion: Fas-mediated apoptosis was cell cycle-specific and initiated at G1 phase.Based on the modified APl and common AnnexinV/PI methods,the establishment of stable and typical cell cycle-specific model in Fas-mediated apoptosis in vitro was feasible.