凋亡相关蛋白的表达及Caspase-3活性改变与人卵巢癌细胞顺铂耐药的关系  

Relationship between Expression of Apoptosis Associated Proteins and Caspase-3 Activity in Cisplatin-Resistant Human Ovarian Cancer Cell Line

作　　者：杨晓葵[1] 郑芳[2] 陈建华[3] 高庆蕾[1] 卢运萍[1] 王世宣[1] 王常玉[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 [2]华中科技大学同济医学院免疫学教研室,湖北武汉430030 [3]武汉大学人民医院妇产科,湖北武汉430060

出　　处：《癌症》2002年第12期1288-1291,共4页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.39870765;39840009);国家杰出青年科学基金项目(No.30025017)

摘　　要：背景与目的:以顺铂为基础的化疗是卵巢癌治疗的重要组成部分,对顺铂的耐药是卵巢癌治疗失败的原因之一。本研究探讨人卵巢癌顺铂耐药细胞株中凋亡相关蛋白表达及caspase-3活性与人卵巢癌细胞顺铂耐药的关系。方法:采用Westernblot法分析人卵巢癌顺铂敏感细胞株COC1和顺铂耐药株COC1/DDP中凋亡相关蛋白bcl-2、bcl-xL、bax、bcl-xS的表达,caspase-3活性和其底物多聚ADP核糖聚合酶(PARP)的变化,并应用流式细胞仪检查不同浓度顺铂作用COC1和COC1/DDP细胞后的细胞凋亡率。结果:在COC1/DDP细胞中,bcl-2和bcl-xL的表达明显高于COC1细胞,bax的表达无明显改变,bcl-xS在COC1和COC1/DDP细胞中均无表达。用顺铂处理后,COC1/DDP细胞中caspase-3活性、PARP裂解片段和凋亡率较COC1细胞均明显降低(P<0.05),且呈浓度依赖性。结论:人卵巢癌细胞对顺铂产生耐药可能与肿瘤细胞内凋亡抑制蛋白过度表达、caspase-3活性下降有关,而与凋亡诱导蛋白的表达无关。Background & Objective:Cisplatin based chemotherapy is an important way for treatment of ovarian cancer, but resistance to cisplatin is one of the reasons of treatment failure of ovarian cancer. This study was designed to investigate the relationship between apoptosis associated proteins and caspase 3 activity as well as their effects on chemoresistance in human ovarian cancer cell lines. Methods:The expression of apoptosis associated proteins (bcl 2,bcl xL,bax,bcl xS),the activity of caspase 3 and cleavage of poly ADP ribose polymerase (PARP) were determined with Western blot analysis in the cisplatin resistant cell (COC1/DDP) and cisplatin sensitive human ovarian cancer cell (COC1). The apoptotic ratios of COC1 and COC1/DDP were measured with flow cytometry after treated with different concentration of cisplatin. Results:The expression of bcl 2 and bcl xL in COC1/DDP cell was significantly higher than that in COC1 cell, whereas the expression of bax showed no change in COC1/DDP and COC1. There was no bcl xS expression in both COC1 and COC1/DDP cells. The activity of caspase 3,the amount of PARP fragments, and apoptotic ratio in COC1/DDP reduced much more than COC1 did after treated with cisplatin. Conclusions:Cisplatin resistance in human ovarian cancer cell lines may associated with the overexpression of anti apoptotic protein bcl 2 and downregulation of caspase 3 activity,but not associated with the expression of bax and bcl xS.

关 键 词：卵巢癌 凋亡相关蛋白 半胱天冬氨酸蛋白酶-3 顺铂 耐药性 化疗 

分 类 号：R737.31[医药卫生—肿瘤]