[Small interference RNA targeting vascular endothelial growth factor inhibits rat corneal neovascularization].
OBJECTIVE
To test the effects of VEGF-siRNA-transfected corneal epithelium on corneal neovascularization (CNV).


METHODS
It was an experimental study. Cultured rat corneal epithelial cells and keratocytes were transfected with synthesize VEGF siRNA by lipofectamine 2000. The level of VEGF mRNA was analyzed by real time PCR, and the protein levels were determined by enzyme-linked immunosorbent assay (ELISA). CNV was induced by cauterization with 1 mol/L sodium hydroxide in rat corneas. The VEGF-siRNA-transfected-corneal epithelium cells were transplanted to the CNV lesions. Immediately after transplantation, the VEGF-siRNA combined with lipofectamine 2000 were directly transfected rat cornea through injecting into the anterior chamber. After surgery, the surface areas occupied by new vessels were measured, and VEGF protein was localized by immunohistochemistry.


RESULTS
The levels of VEGF expression at both mRNA and protein in the VEGF-siRNA transfected corneal epithelial cells and keratocytes were significantly lower than those of control cells. VEGF siRNA could inhibit the expression of VEGF mRNA in corneal epithelial cells and keratocytes to 57% - 85% and 59% - 78%, respectively. The VEGF-siRNA-transfected-corneal epithelium transplantation significantly decreased the surface areas occupied by new vessels. VEGF expression level in interference groups was lower than that in the control group.


CONCLUSIONS
The development of CNV is markedly suppressed by VEGF-siRNA transfection in vivo.
Fan
Xian-qun
XQ
Department of Ophthalmology, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China. fanxq@sh163.net
Li
Jin
J
Fu
Yao
Y
Jia
Ren-bing
RB
Lu
Wen-juan
WJ
D000818:Animals; D002478:Cells, Cultured; D016510:Corneal Neovascularization / Q000235:genetics* / Q000378:metabolism; D019573:Epithelium, Corneal / Q000378:metabolism; D015316:Genetic Therapy; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors*
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A