Effect of 17-AAG, a Hsp90 inhibitor, on the expression of ILK in RPE cells under hypoxic condition

Abstract:

AIM: To investigate the effect of 17-allylamio-17-demethoxygeldanamycin(17-AAG), an heat shock protein 90(Hsp90)inhibitor, on the expression of integrin- linked kinase(ILK)induced by hypoxia in retinal pigment epithelium(RPE)cells.

METHODS: RPE cells were cultured with 200μmol/L cobalt chloride(CoCl₂)for 12 hours to imitate chemical hypoxia. Pretreatment of 17-AAG was 1 hour prior to hypoxic insult. Experiment was divided into hypoxic control group, Dimethyl sulfoxide(DMSO)control group and 17-AAG pretreatment group(0.01, 0.10, 0.50, 1.00, 5.00 and 10.00μmol/L). RT-PCR and Western blot analysis were used to examine the expression of ILK in cultured RPE cells.

RESULTS: The density ratios of ILK mRNA and β-actin mRNA of hypoxic control group, DMSO control group and 17-AAG pretreatment groups were 1.32±0.04,1.29±0.03,0.93±0.06,0.70±0.05,0.53±0.03,0.44±0.04,0.32±0.04,0.30±0.03; and the density ratios of ILK protein and β-actin protein were 2.16±0.04, 2.13±0.04, 1.65±0.04, 1.13±0.05, 0.74±0.03, 0.41±0.06, 0.35±0.04, 0.35±0.03. The ILK expression in 17-AAG pretreated groups were inhibited compared to hypoxic control group(P < 0.05), and the decrease was in concentration-dependent manner.

CONCLUSION: Hsp90 inhibitor 17-AAG was effective to inhibit the expression of ILK induced by hypoxia in RPE cells.