拓扑替康诱导卵巢癌COC1/DDP细胞凋亡的分子机制初步探讨  

Molecular Mechanism of Apoptosis in Cisplatin-resistant Human Ovarian Cancer Cell Line Promoted by Topotecan

作　　者：田训[1] 王薇[1] 邢辉[1] 徐茜[1] 李辅军[1] 卢运萍[1] 王世宣[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《肿瘤防治研究》2005年第6期360-363,共4页Cancer Research on Prevention and Treatment

基　　金：国家重点基础研究发展项目(2002CB513100)

摘　　要：目的探讨拓扑替康(TPT)对人卵巢癌顺铂耐药细胞株COC1/DDP的杀伤和诱导凋亡活性及其作用机制。方法MTT比色法与软琼脂克隆形成测定TPT对人卵巢癌顺铂耐药细胞株COC1/DDP的杀伤效应;TUNEL法和流式细胞仪研究TPT对靶细胞的凋亡诱导作用;Westernblot检测TPT对COC1/DDP细胞内bcl-2、bax和caspase-3基因表达的影响以及caspase-3活性的改变。结果TPT对COC1/DDP细胞有明显细胞毒性作用,不仅有剂量依赖性,也存在明显的时间依赖性;COC1/DDP细胞在TPT作用后出现特征性凋亡形态特征,且凋亡率由8.54%上升为23.16%(P<0.05)。TPT不影响COC1/DDP细胞内bcl-2蛋白表达,却明显增加bax蛋白和caspase-3蛋白表达,并提高caspase-3活性。结论TPT对人卵巢癌顺铂耐药细胞株COC1/DDP有明显的杀伤和促凋亡作用,其机制可能依赖于细胞内bax蛋白表达增高和caspase-3的活化。Objective To investigate the cytotoxicity and proapoptotic activity of topotecan(TPT) on cisplatin-resistant human ovarian cancer cell lines COC1/DDP and its mocular mechanisms.Methods The cytotoxicity of TPT on COC1/DDP was monitered using MTT assay and soft agaragar assay. TUNEL and flow cytometry were employed to observe the apoptosis of COC1/DDP induced by topotecan. Western blot were used to determine the expression of bcl-2, bax and caspase-3, in addition to the activity of caspase-3.Results TPT had strong cytotoxicity on COC1/DDP cell in vitro, and the cytotoxicity was dose-and time-dependent. TPT-treated cells demonstrated apoptosis property while the apoptotic rate was increased from 8.54% to 23.16% ( P < 0.05 ). Although the expression of bcl-2 was not affected, the expression of bax and caspase-3 were upregulated and the caspase-3 activity was promoted in the course of COC1/DDP apoptosis when treated with topotecan. Conclusion The cytotoxicity and proapoptosis activity of TPT, which is significantly observed in cisplatin- resistant human ovarian cancer, may be related to the increased bax expression and the promoted caspase-3 activity.

关 键 词：拓扑替康 卵巢癌 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]