[Inhibition of vegfr-2 gene expression by RNA interference].
This study was aimed to explore a novel potential gene target for therapy of malignant tumor. The recombinant expression plasmids of VEGF/VEGFR-2 were designed, constructed and then transfected into A549 cells by using lipofectamine. The expressions of VEGF/VEGFR-2 mRNA and protein were detected by RT-PCR and Western blot respectively. The cell proliferation was assayed by CCK-8 and the cell apoptosis was detected using Hoechst Staining. The results indicated that as compared with the blank control, pGenesil-1 and scramble groups, the interference effect of pGenesil-1-vegfr-2-shRNA-1 vector group was more obvious. As the expression of endogenous vegfr-2 mRNA decreased, the expression of VEGFR-2 protein decreased correspondingly. The proliferation of A549 cells was inhibited significantly by RNAi at 72 hours (p<0.01). The apoptosis of A549 cells was induced at 48 hours after being transfected with pGenesil-1-vegfr-2-shRNA-1 and the typical apoptosis morphology could be seen by fluorescence microscopy. It is concluded that the expression of vegfr-2 gene is inhibited effectively by vegfr-2 specific shRNA. The proliferation of A549 cells is inhibited significantly and the apoptosis of cells is induced. This result showed that VEGF/VEGFR-2 signaling pathway can be an effective target for the prevention and treatment of malignant tumor.
Department of Pediatrics, Guangdong Medical College Affiliated Hospital, Zhanjiang 524023, Guangdong Province, China. xiaodan830302@126.com
Xiao-Dan
XD
Chang
Ming-Zhen
MZ
Chen
Zhong-Lu
ZL
Ye
D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics*
D034741:RNA, Small Interfering; D040301:Vascular Endothelial Growth Factor Receptor-2