Inhibitory Effects Of Chemically Synthetic Sirna Of HIF-1α In Rat Retinal Endothelial Cells Of Hypoxic Condition

Objective To study blocking effects of RNAi technology on HIF-1αexpression in rat retinal endothelial cells.Methods After liposome-mediated siRNA transfection of rat retinalendothelial cells，the cells were cultured in CoCl2-induced hypoxiccondition for8hours，the expression of HIF-1αmRNA was determined byfluorenscence quantitative PCR， HIF-1α protein expression was detectedby Western Blot. Cell proliferation was measured with MTT assay.Results Rat retinal endothelial cells were successfully transfectedwith siRNA. Fluorescence quantitative PCR results showed that aftertransfected48hours，the interference group of siRNA1and siRNA2on theexpression of HIF-1α gene was significantly reduced(t=16.786,8.953,4.087，P＜0.05)，Western Blot result showed that HIF-1α protein was significantly inhibited in interference group of siRNA1and siRNA2(t=6.861,2.893,4.567,5.072， P＜0.05). Decreased celluarproliferation activity was observed by MTT，and49.50%and67.41%of cellsgrowth inhibition was observed after transfecting with siRNA1and siRNA2respectively.Conclusion The synthetic HIF-1αsiRNA can effectively inhibit theexpression of HIF-1α gene and reduce cell proliferation in rat retinalendothelial cells of hypoxic condition. RNA interference technology withHIF-1α as target is expected to become a new strategy for gene therapyof ROP.