[Effects of hemin and thermal stress exposure on JWA expression].
OBJECTIVE
To investigate the expression of JWA after hemin and (or) thermal stress exposure.


METHODS
The expression of JWA was determined by Western blot. RT-PCR was carried out for evaluation of the expression of JWA mRNA. The JWA promoter transcription activity analysis was performed by CAT-ELISA.


RESULTS
The expression of JWA protein was significantly increased by 3.23 +/- 0.57 times of control in K562 cells after treated by hemin (50 micromol/L) for 1 week, and the similar pattern was observed in the cells after treatment with thermal stress (42 degrees C) for 2 hours (increased by 8.00 +/- 1.73 times). The expression of JWA mRNA was also significantly elevated by 1.37 +/- 0.06 times in K562 cells treated by hemin (30 micromol/L) for 48 hours, and with a similar regulation pattern (increased by 1.87 +/- 0.13 times) by treatment with thermal stress (42 degrees C) for 30 minutes. However, an antagonistic effect was observed by treatment of K562 cells with hemin (30 micromol/L, 48 hours) followed by thermal stress (42 degrees C, 30 minutes). The CAT-ELISA further confirmed that hemin or thermal stress treatment alone up-regulated JWA transcription activity while the effects could be counteracted partly by the combined treatment of the both.


CONCLUSIONS
The hemin and thermal stress may regulate JWA expression via distinct intracellular signal transduction pathways.
Zhao
Ming
M
Department of Molecular Cell Biology & Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, China.
Chen
Rui
R
Li
Ai-ping
AP
Zhou
Jian-wei
JW
D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D018882:Heat Stress Disorders / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006427:Hemin / Q000494:pharmacology*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D020014:K562 Cells; D026901:Membrane Transport Proteins; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
C484566:ARL6IP5 protein, human; D006360:Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D006427:Hemin