Zhu Ting, Li Ai-ping, Liu Qi-zhan, Wang Xin-ru, Zhou Jian-wei
Department of Molecular Cell Biology and Toxicology, Jiangsu Provincial Key Laboratories of Human Functional Genomics and of Toxicology, Nanjing Medical University, Nanjing 210029, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2005 May;39(3):182-6.
To investigate the expression of JWA gene, heat shock proteins (hsp70 and hsp27) and p53, and to explore the role and the possible mechanism of JWA gene involved in H2O2-induced oxidative stress of K562 cells.
0.01, 0.1, 1 mmol/L H2O2 treated K562 cells at 10, 30, 60 and 180 min to established the models of DNA damage. Furthermore, K562 cells were induced apoptosis by 0.1 mmol/L H2O2 at different time (6-48 h) and different concentration (0.5-1,000 micromol/L) of H2O2 at 48 h. DNA damage and cell apoptosis were detected by DNA gel electrophoresis. And the immunoblotting assay was used for detecting expressions of JWA protein and correlated genes (hsp27, hsp70 and p53).
During the DNA damage, JWA was much more sensitive to H2O2 than those heat shock proteins, and its expression pattern was very similar to that of hsp70. And at low concentration of H2O2-exposure (0.01 mmol/L), the expressions of JWA and heat shock proteins were all increased greatly. In addition, JWA, hsp70, hsp27 and p53 overexpressed and their expression pattern were similar during cell apoptosis.
JWA should be functioning as an effective environmental responsive gene and should actively participate the signal pathways of oxidative stress which might be associated with hsp70 and p53.
研究JWA基因、热休克蛋白(hsp70和hsp27)及p53的表达情况,探讨JWA基因在H2O2诱导的K562细胞氧化应激中的作用及可能机制。
用0.01、0.1、1 mmol/L H2O2处理K562细胞10、30、60和180分钟,建立DNA损伤模型。此外,用0.1 mmol/L H2O2在不同时间(6 - 48小时)以及在48小时用不同浓度(0.5 - 1000微摩尔/升)的H2O2诱导K562细胞凋亡。通过DNA凝胶电泳检测DNA损伤和细胞凋亡情况。采用免疫印迹法检测JWA蛋白及相关基因(hsp27、hsp70和p53)的表达。
在DNA损伤过程中,JWA对H2O2比热休克蛋白更敏感,其表达模式与hsp70非常相似。在低浓度H2O2暴露(0.01 mmol/L)时,JWA和热休克蛋白的表达均大幅增加。此外,在细胞凋亡过程中,JWA、hsp70、hsp27和p53均过表达,且表达模式相似。
JWA应作为一种有效的环境响应基因,积极参与氧化应激信号通路,可能与hsp70和p53有关。