JWA基因参与细胞氧化应激的机制

[The mechanism of JWA gene involved in oxidative stress of cells].

作者信息

Wang Nan-ping, Zhou Jian-wei, Li Ai-ping, Cao Hai-xia, Wang Xin-ru

机构信息

Department of Preventive Medicine, Medical College, Three Gorges University, Yichang 443003, China.

出版信息

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2003 Jun;21(3):212-5.

PMID:14761492

Abstract

OBJECTIVE

To study the mechanism of gene JWA involved in oxidative stress under hydrogen peroxide (H(2)O(2)) exposure.

METHODS

Both MCF-7 (human breast cancer cell line) and WI-38 (human embryo lung fibroblast cell line) cells were treated with 1 mmol/L of H(2)O(2) with or without pre-incubation of taurine (tau). Malondialdehyde (MDA) and glutathione (GSH) contents in supernatant of cell culture were measured; RT-PCR and Western blotting were carried out for evaluation of the expressions of JWA mRNA and protein respectively. Heat shock proteins (HSP27, HSP70 and HSP90) were also analyzed.

RESULTS

The contents of MDA before and after H(2)O(2) treatment in MCF-7 cells were (0.531 +/- 0.038), (0.674 +/- 0.410) mmol/L respectively, (P < 0.01), while those in WI-38 cells were (0.572 +/- 0.035), (0.683 +/- 0.028) mmol/L respectively, (P < 0.01). The contents of GSH before and after H(2)O(2) treatment in MCF-7 cells were (0.053 +/- 0.002), (0.044 +/- 0.002) g/L respectively, (P < 0.01), while those in WI-38 cells were (0.058 +/- 0.002), (0.050 +/- 0.002) g/L respectively, (P < 0.01). The expression of JWA mRNA was down regulated, at 6 h it decreased by 68.4%, while in WI-38 cells no obvious change found. JWA protein and HSP27 showed markedly increased after H(2)O(2) treatment in both cells but not in similar extent.

CONCLUSION

Oxidative stress signal pathways of JWA gene varied between cancer and non-cancer cell lines; JWA protein may have a similar function as HSP27 and act as an important signal molecule in H(2)O(2) induced cell injury.

摘要

目的

研究基因JWA在过氧化氢（H₂O₂）暴露下参与氧化应激的机制。

方法

用1 mmol/L的H₂O₂处理MCF-7（人乳腺癌细胞系）和WI-38（人胚肺成纤维细胞系）细胞，同时设置有无预先孵育牛磺酸（tau）的组。检测细胞培养上清液中丙二醛（MDA）和谷胱甘肽（GSH）的含量；分别采用RT-PCR和蛋白质印迹法评估JWA mRNA和蛋白的表达。同时分析热休克蛋白（HSP27、HSP70和HSP90）。

结果

MCF-7细胞在H₂O₂处理前后MDA含量分别为（0.531±0.038）、（0.674±0.410）mmol/L，（P<0.01）；WI-38细胞分别为（0.572±0.035）、（0.683±0.028）mmol/L，（P<0.01）。MCF-7细胞在H₂O₂处理前后GSH含量分别为（0.053±0.002）、（0.044±0.002）g/L，（P<0.01）；WI-38细胞分别为（0.058±0.002）、（0.050±0.002）g/L，（P<0.01）。JWA mRNA表达下调，6小时时下降了68.4%，而WI-38细胞中未发现明显变化。H₂O₂处理后，两种细胞中JWA蛋白和HSP27均显著增加，但程度不同。