新的环境应答蛋白JWA在MCF-7细胞应答氧化应激中的表达  

Expression of novel environmental responsive protein JWA involved in the oxidative stress responsiveness in MCF-7 cells

作　　者：朱婷[1] 陈瑞[1] 李爱萍[1] 顾灯安[1] 刘起展[1] 周建伟[1] 

机构地区：[1]南京医科大学公共卫生学院江苏省人类功能基因组学和应用毒理学重点实验室分子毒理研究室,210029

出　　处：《中华劳动卫生职业病杂志》2005年第2期122-124,i002,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目(30170812);国家"973"重大基础研究项目(2002CB512900);国家科技部重大研究项目前期专项资助项目(2001 50)

摘　　要：目的　研究氧化应激诱导MCF 7 细胞氧化损伤过程中,JWA表达的调节规律及其作用,尤其是与热休克蛋白(HSPs)间的关系。方法　以不同浓度的H2O2(0.01、0.10、1.00 mmol/L)分别处理MCF 7细胞10、30、60、180 min,建立MCF 7细胞氧化损伤模型并用DNA琼脂糖凝胶电泳鉴定;用四唑蓝(MTT)法分析H2O2 对MCF 7细胞的相对细胞增殖率和细胞毒性的影响;采用Western blot方法检测JWA、HSP70、HSP27和热休克转录因子(HSF1)蛋白表达水平的变化。结果　H2O2 对MCF 7细胞的增殖抑制和细胞毒性呈时间和剂量依赖关系,MCF 7细胞在最高剂量、最长处理时间(1.00 mmol/L、180 min)时,细胞增殖几乎完全受抑制。H2O2 活跃地调节JWA的表达,氧化损伤使JWA、HSP70 和HSF1表达上调并呈剂量依赖关系,而且JWA、HSP70和HSF1的表达规律相似,而HSP27 表达似乎与氧化应激的信号调节无关。结论　在H2O2 致MCF 7 细胞氧化应激时,JWA作为有效的环境应答蛋白,其与HSP70共同参与的信号通路可能受HSF1调控。Objective To study the expression and the possible role of JWA protein in oxidative stress-induced damage of MCF-7 cells,especially the relationship between JWA and heat shock proteins(HSPs). Methods MCF-7 cells were exposed to different concentration of H 2O 2(0.01,0.10,1.00 mmol/L) for different time(10,30,60 and 180 min) respectively.DNA damage was detected by using DNA gel electrophoresis.The MTT assay was used to analyze the effect of H 2O 2 on the cytotoxicity and relative cell proliferation ratio of the cells.The expressions of JWA,HSP70,HSP27 and HSF1 were determined by Western-blot. Results The inhibitory effect on MCF-7 cells viability induced by H 2O 2 was shown a dose-and time-dependent manner and MCF-7 cells proliferation,and was almost completely inhibited by the exposure of H 2O 2 at 1.00 mmol/L for 180 min.Hydrogen peroxide treatment of MCF-7 cells caused oxidative stress which up-regulated the expressions of JWA,HSP70 and heat shock factor 1(HSF1) in a dose-dependent manner,and the expression pattern of JWA was very similar to those of HSP70 and HSF1 but not to HSP27. Conclusion JWA might enhance intracellular defenses against H 2O 2-induced oxidative damage in human breast carcinoma cells.JWA is determined functioning as an effective environmental responsive protein and as a parallel molecule of HSP70 actively participates in the signal pathways of oxidative damage which might be regulated by HSF1.

关 键 词：JWA基因 氧化性应激 热休克蛋白质类 肿瘤 发生机制 

分 类 号：R73-3[医药卫生—肿瘤]