[Effect of vascular endothelial growth factor small interfering RNA (siRNA) on retinal microvascular endothelial cells under hypoxia condition in vitro].
OBJECTIVE
To explore VEGF siRNA's effect on the immature fetal retinal microvascular endothelial cells in vitro.


METHODS
The fresh retinal micrangium was primarily cultured to obtain microvascular endothelial cells. CoCl2 was used to simulate oxygen-deficient conditions. siRNA directed against human VEGF was designed and chemically synthesized. There were 3 groups in our experiment: VEGF siRNA group, hypoxia control group, and negative siRNA control group. The fetal retinal micrangium vascular endothelial cells were transfected by using liposome. The expression levels of VEGF mRNA and protein were evaluated by RT-PCR and Western blotting 24, 48, 72 h after transfection, cell proliferation was evaluated by MTT method.


RESULTS
The expression levels of VEGF mRNA decreased by 21.05%, 79.67%, and 90.48% 24 h, 48 h, and 72 h after transfection as compared to those in hypoxia control group, the expression level of VEGF protein had decreased by 14.58%, 66.97%, and 81.61% as compared to those in hypoxia control group. The siRNA could decrease cell proliferation under hypoxia too, the multiplication rate after 12, 24, 48, and 72 h decreased by 15.0%, 42.9%, 78.3% and 65.9%.


CONCLUSIONS
VEGF siRNA could down-regulate the expression of VEGF in immature fetal retinal microvascular endothelial cells and suppressed cell proliferation. Application of siRNA to inhibit expression of VEGF may be a hopeful way to prevent and cure ROP.
Zhang
Xiao-zhuang
XZ
Department of Pediatrics, Guangdong Province Maternal and Childern's Hospital, Guangzhou 510010, China.
Jiang
Jing-bo
JB
Luo
Xian-qiong
XQ
Huang
Shui-qing
SQ
Yang
Jie
J
Li
Jian-qiao
JQ
Lin
Qiu-xiong
QX
D015687:Cell Hypoxia; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D007231:Infant, Newborn; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D012160:Retina / Q000378:metabolism / Q000473:pathology; D012171:Retinal Vessels / Q000166:cytology* / Q000378:metabolism; D012178:Retinopathy of Prematurity / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger; D034741:RNA, Small Interfering; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A