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Inhibiting The Protein Expression Of Vascular Endothelial Growth Factor (VEGF) In Human Tongue Carcinoma Cell By Small Interfering RNA Targeting VEGF

Posted on:2007-12-12Degree:MasterType:Thesis
Country:ChinaCandidate:Z W YaoFull Text:PDF
GTID:2144360185952276Subject:Oral and Maxillofacial Surgery
Abstract/Summary:PDF Full Text Request
Purpose (1)Construct VEGF SiRNA vector. (2)examine the effectiveness on the specific inhibition of VEGF expression in transfected human Tongue carcinoma cell (Tca8113) by SiRNA.Methods (1)Two pairs of SiRNA sequences were synthesized. With the plasmid Psilencer2.1-U6-neo, VEGF SiRNA vectors were successfully constructed, which were named Psilencer2.1-U6-neo /SiRNA1 and Psilencer2.1-U6-neo /SiRNA2 respectively. (2) Psilencer2.1-U6-neo /SiRNAl and Psilencer2.1-U6-neo /SiRNA2 and control plasmid were transfected into Tca8113 cells by cationic polymer mediated method. The transfected cells were selected with 400mg/l G418 for 8 d . Clones were picked and expanded. The Tca8113 cells in normal culture medium was taken as negative control group. The expression of VEGF protein in tansfected Tca8113 cells in each group was staining and measured by Immuno-cytochemical method, and Medical colour-image measure system for Immuno- cytochemistry was used for The expression of VEGF protein, which results was described by gray scale. The concentration of the VEGF protein in culture medium secreted by transfected and control Tca8113 cells was measured by ELISA kits.Results (1)Vector of VEGF SiRNA was constructed successfully; (2)VEGF SiRNA1 and SiRNA2 expressing vectors were transfected into Tca8113 cells. The relative amount of VEGF protein in cells in Psilencer2.1-U6-neo /SiRNA1,Psilencer2.1-U6-neo /SiRNA2, control...
Keywords/Search Tags:small interfering RNAs,SiRNAs, vascular endothelial growth factor,VEGF, tongue carcinoma
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