[Expression of novel environmental responsive protein JWA involved in the oxidative stress responsiveness in MCF-7 cells].
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(2)O(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(2)O(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(2)O(2) was shown a dose-and time-dependent manner and MCF-7 cells proliferation, and was almost completely inhibited by the exposure of H(2)O(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.


CONCLUSIONS
JWA might enhance intracellular defenses against H(2)O(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.
Zhu
Ting
T
Department of Molecular Cell Biology & Toxicology, Jiangsu Provincial Key Laboratories of Human Functional Genomics and of Applied Toxicology, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu Province 210029, China.
Chen
Rui
R
Li
Ai-ping
AP
Gu
Deng-an
DA
Liu
Qi-zhan
QZ
Zhou
Jian-wei
JW
D045744:Cell Line, Tumor; D004249:DNA Damage / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005260:Female; D055551:HSP27 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D006801:Humans; D006861:Hydrogen Peroxide / Q000009:adverse effects*; D047908:Intracellular Signaling Peptides and Proteins; D026901:Membrane Transport Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000096:biosynthesis; D018384:Oxidative Stress / Q000187:drug effects*; D014157:Transcription Factors / Q000096:biosynthesis; D015854:Up-Regulation
C484566:ARL6IP5 protein, human; D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D055551:HSP27 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; C057725:HSPB1 protein, human; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D026901:Membrane Transport Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D014157:Transcription Factors; D006861:Hydrogen Peroxide