17-AAG, a Hsp90 inhibitor, attenuates the hypoxia-induced expression of SDF-1alpha and ILK in mouse RPE cells.
The aim of this study was to investigate the changes of SDF-1alpha and ILK expression in mouse retinal pigment epithelium (RPE) cells in response to hypoxia, and the effect of 17-Allylamino-17-demethoxygeldanamycin (17-AAG), a heat shock protein 90 (Hsp90) inhibitor, on the hypoxia-induced expression of SDF-1alpha and ILK. RPE cells were cultured with 200 micromol/L cobalt chloride (CoCl(2)) for different times (1, 3, 6, 12, 24, 72 h) to imitate chemical hypoxia. Pretreatment of 17-AAG was 1 h prior to hypoxic insult. Cellular viability after 17-AAG treatment was assessed by MTT assay, and the changes of SDF-1alpha and ILK expression were examined by RT-PCR and Western blot. Up-regulation of SDF-1alpha and ILK expression in response to hypoxia was observed. One hour pretreatment of 17-AAG could remarkably decreased the hypoxia-induced SDF-1alpha and ILK expression in vitro. Our results indicated that SDF-1alpha and ILK involved in the hypoxic response of RPE cells, and 1 h pretreatment of 17-AAG had an inhibitive effect on the hypoxia-induced SDF-1alpha and ILK expression.
Wang
Ye Qing
YQ
Ophthalmology Hospital, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Zhang
Xiao Mei
XM
Wang
Xiao Dan
XD
Wang
Bin Jie
BJ
Wang
Wei
W
D000818:Animals; D016227:Benzoquinones / Q000494:pharmacology*; D015153:Blotting, Western; D015687:Cell Hypoxia / Q000187:drug effects; D002460:Cell Line; D054377:Chemokine CXCL12 / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors*; D047029:Lactams, Macrocyclic / Q000494:pharmacology*; D051379:Mice; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D055213:Retinal Pigment Epithelium / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000099200:Scaffold Protein ILK
D016227:Benzoquinones; D054377:Chemokine CXCL12; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D017346:Protein Serine-Threonine Kinases; D000099200:Scaffold Protein ILK; C112765:tanespimycin