拓扑替康促人卵巢癌顺铂耐药细胞株凋亡机制的初步研究  

Molecular mechanism of apoptosis in cisplatin-resistant human ovarian cancer cell lines promoted by topotecan

作　　者：王薇[1] 邬素芳[1] 高庆蕾[1] 杨晓葵[1] 奚玲[1] 王世宣[1] 卢运萍[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

出　　处：《肿瘤》2004年第6期554-557,共4页Tumor

基　　金：国家重点基础研究发展项目 (编号 :2 0 0 2CB5 13 10 0 );国家杰出青年科学基金资助项目 (编号 :3 0 0 2 5 0 17)

摘　　要：目的　探讨拓扑替康 (TPT)对人卵巢癌顺铂耐药细胞株A2 780 /DDP和COC1/DDP的杀伤和诱导凋亡活性及其作用机制。方法　用MTT比色法测定TPT对人卵巢癌顺铂耐药细胞株A2 780 /DDP和COC1/DDP的杀伤效应 ;透射电镜和DNA凝胶电泳研究TPT对靶细胞的凋亡诱导活性 ;Westernblot检测凋亡相关蛋白bcl 2和bax的表达。结果　TPT对A2 780 /DDP和COC1/DDP细胞均有较强的体外杀伤作用 ,其IC50 值分别为 10 2 .91ng/ml和 111.75ng/ml;TPT能诱导A2 780 /DDP和COC1/DDP细胞产生凋亡 ,DNA凝胶电泳呈现典型的凋亡梯度 ,透射电镜观察到细胞核染色质固缩、边集 ,胞质浓缩 ,出现空泡等典型的凋亡超微结构改变 ;TPT不影响bcl 2蛋白表达 ,却能上调bax蛋白表达 ,增大bax/bcl 2比值 ,且呈剂量依赖性。结论　TPT对人卵巢癌顺铂耐药细胞株A2 780 /DDP和COC1/DDP均有明显的杀伤和促凋亡作用 ,其机制可能与bax基因表达上调而提高bax/bclObjective To investigate the cytotoxicity and apoptotic activity of topotecan(TPT) on cisplatin-resistant human ovarian cancer cell lines A2780/DDP and COC1/DDP.Methods The cytotoxicity of TPT on A2780/DDP and COC1/DDP were monitored by MTT assay.TPT-induced apoptosis was determined by DNA ladder and transmission electron microscope(TEM).Western blot was employed to observe the expression of bcl-2 and bax. Results TPT had strong cytotoxicity on A2780/DDP and COC1/DDP cells in vitro and the drug IC 50 were 102.91ng/ml and 111.75ng/ml, respectively.TPT-treated cells demonstrated apoptosis property such as typical DNA ladder,nuclear chromatin pycnosis and margination,cell shrinkage, and cytoplasm condensation.Western blot showed that bcl-2 expression was not fluctuated,but the level of bax expression was upregulated with a dose-dependent pattern,The bax/bcl-2 ratio went up when TPT dose increased over 50ng/ml. Conclusion The cytotoxicity and proapoptosis activity of TPT, is significantly observed in cisplatin- resistant human ovarian cancer,cell line may be related to high expression of bax and the increasing of bax/bcl -2.

关 键 词：卵巢肿瘤 拓扑替康 细胞凋亡 BCL-2蛋白 BAX蛋白 

分 类 号：R730.53[医药卫生—肿瘤]