Selection of the most effective small interfering RNA which inhibits the expression of caveolin-1 in endothelial cells and detection of the cytotoxicity of transfection complex

Objective Aim to pick out the small interference RNA (siRNA) which could most effectively inhibit the expression of caveolin-1 in endothelial cells and to detect the cytotoxicity of the transfection complex.Methods Four siRNAs were chemically synthesized:siRNA422,siRNA548,siRNA710 were used to inhibit caveolin-1 expression in endothelial cells,FAM-labeled mismatch siRNA was as a negative control They were all transfected into endothelial cells,respectively.Caveolin-1 mRNA was detected after transfection by real-time PCR and its protein expression was measured by Western blot.The cytotoxicity was analyzed with the method of the sulforhodamine B (SRB).Results ① The transfecfon rates are all above 85％ under the transfection condition of siRNA(20,30 nmol/L or 40 nmol/L) combined with the same dose of siRNAFect(P＜0.01).② The survival rates of transfected cells are all above 80％ under the transfection condition of siRNAFect 1.3 μl combined with siRNA (10 nmol/L or 20 nmol/L)(P＜0.05).③ The expression of caveolin-1 mRNA in EA.hy926 cells was significantly decreased after siRNA548 treatment (P＜0.01).④ Compared with siRNA422,siRNA710,vehicle or mismatch siRNA-treated group,the expression of caveolin-1 protein was the least after siRNA548 treatment 48 h later(P＜0.01).Conclusions ① The siRNA548 can inhibit caveolin-1 expression most effectively in endothelial cells.② siRNA(20 nmol/L) combined with siRNAFect(1.3 μl) has lower cytotoxicity and higher transfection efficiency,which is suitable for transfection. Key words: Endothelial cells;  Caveolin-1 ;  Small interfering RNA;  Sulforhodamine B;  Cytotoxicity;  Election