RNA interference can block VEGF upregulation in hypoxic corneal epithelial cells

Abstract

Abstract: : Purpose: To determine if RNA interference could block hypoxia–induced upregulation of VEGF in corneal epithelial cells. Methods: Human corneal epithelial cells were transfected with siRNA expression cassettes targeting VEGF or a nonspecific sequence. Two days later, they were subjected to 5% hypoxia for 48h. Supernatant was sampled at 24 and 48 h and assayed for VEGF by ELISA. Results: Baseline VEGF concentration was 528 pg/mL; this increased to 794 and 645 pg/mL at 24 and 48h of hypoxia, increases of 76% and 43%, respectively.Two siRNA sequences targeting VEGF were able to limit VEGF increases to 13% and 12% at 24h, and resulted in –59% and 14% changes at 48h (all p's<0.05). Nonspecific siRNA sequences had no significant effect. Conclusions: RNA interference can significantly suppress expression of VEGF induced by hypoxia in corneal epithelial cells.

Keywords: cornea: epithelium • neovascularization • hypoxia