manumycin诱导舌鳞癌Tca8113细胞凋亡的作用及机制  

Effects and mechanism of manumycin induced apoptosis of Tca8113 cells

作　　者：王葆[1] 梁永钜[1] 符立梧[1] 许光普[1] 

机构地区：[1]华南肿瘤学国家重点实验室

出　　处：《中国药理学通报》2006年第9期1104-1111,共8页Chinese Pharmacological Bulletin

摘　　要：目的研究manumycin抑制Tca8113细胞的生长和诱导细胞凋亡的作用及机制。方法细胞毒作用以MTT法测定;凋亡细胞形态以Hoechst33258染色后荧光显微镜观察;凋亡率的检测以AnnexinV染色,流式细胞仪检测;细胞内活性氧(ROS)和线粒体跨膜电位(ΔΨm)分别用DCFH-DA和DiOC6荧光探针标记,流式细胞仪检测;蛋白质定量检测以Westernblot法。结果manumycin浓度依赖性抑制Tca8113细胞的生长,IC50为(11.33±0.63)μmol.L-1;manumycin可浓度和时间依赖性诱导Tca8113细胞的凋亡,过氧化物清除剂N-乙酰半胱氨酸能清除manumycin介导的细胞活性氧的增加,抑制线粒体跨膜电位降低及细胞凋亡。结论manumycin体外显著抑制舌鳞癌Tca8113的生长及诱导细胞凋亡,其机制可能通过激活线粒体依赖性凋亡通路有关。Aim To determine effects and mechanism of manumycin on growth inhibition and inducing apoptosis of Tca8113 cells. Methods Cytotoxicity was determined by tetrazolium （MTT） assay. Apoptotic morphology of Tca8113 cells was observed under fluorescent microscope after Hoechst33258 staining. Annexin V stain was used to examine the apoptosis of Tca8113 cells induced by manumycin. Reactive oxygen species （ROS） levels and mitochondrial membrane potential （△ψm） in cells were labeled with DCFH-DA and DiOC6 and tested by flow cytometry （FCM）. Activation of proteins was measured by Western blot. Results manumycin was shown to inhibit the growth of Tca8113 cells significantly in a dose-dependent manner, with mean IC50 of 11.33 μmol·L^-1. manumycin induced apoptosis of Tca8113 cells in a time-dependent and concentration-dependent manner. The generation of ROS increased obviously after cells were treated with manumycin and the ROS can be cleared by NAC, meanwhile the decrease of △ψm and the apoptosis of Tca8113 cells could be restrained by NAC. Conclusion manumycin showed inhibitory effects on Tca8113 cells and induced apoptosis of Tca8113 cells. The mechanism of the action was associated with the increase in the cellular ROS level and the decrease of the mitochondrial membrane potential induced by manumycin, which resulted in activating chondriosome passway and releasing of casepase activator.

关 键 词：manumycin活性氧 N-乙酰半胱氨酸 线粒体膜电位 细胞凋亡 CASPASE 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]