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.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
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)
