Title: Short Hairpin RNA Targeting Vascular Endothelial Growth Factor Inhibits Expression of Vascular Endothelial Cell in Human GBC-SD Cells
Abstract: Objective:To construct a plasmid expression vector coding for the short hairpin RNA(shRNA) targeting VEGF and to evaluate the effect of VEGF shRNA on expression of VEGF in human GBC-SD cells.Methods:To designe four pairs of shRNAs that target at VEGF gene.To analyses the sequences of the shRNA.The plasmids were then transfected into GBC-SD cells via liposome2000.The VEGF mRNA was determined using reverse transcriptase-polymerase chain reaction.The VEGF protein was determined using enzyme-linked immunosorbent assay and Western blot analyses.Results:The expression plasmids were confirmed by sequence analysis.The VEGF mRNA levels of shRNA2 group the shRNA plasmids resulted in a inhibition of VEGF mRNA by 86%.The protein expression of VEGF was inhibited effectually in VEGF1-4 Groups by enzyme-linked immunosorbent assay and Western blot analyses.There was not obvious change in the NC group.Conclusions:The shRNA eukaryotic expression plasmid targeting at VEGF gene is constructed successfully.The VEGF mRNA and protein was suppressed signifi-cantly by this given plasmid.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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