The Biological Effects And The Mechanism Of Sirna Targeting VEGF Plus Paclitaxel In Endometrial Carcinoma Ishikawa Cell Line

Purpose:To observe the biological effects and the mechanism of SiRNA targeting VEGF plus Paclitaxel in endometrial carcinoma Ishikawa cell line.Method:Small interfering RNA (siRNA) targeting VEGF gene was designed, then ds-siRNA was transfected into Ishikawa cells with LipfectamineTM2000;The effect of different concentrations of Paclitaxel on cellular Proliferation was analyzed by MTT assay; human endometrial carcinoma Ishikawa cells were divided into five different treatment groups:control, siRNA-N, siRNA-VEGF, Paclitaxel, siRNA-VEGF+Paclitaxe, expressions of VEGF on mRNA and Protein level were analyzed by Realtime-PCR and Western blotting respectively, cell cycle and apoptosis were measured by flow cytometry(FCM).Ability of invasion was measured by transwell chamber model.Results:Teatment with siRNA-VEGF、 Paclitaxel、siRNA-VEGF plus with Paclitaxel decreased the expression of VEGF on both mRNA and protein level and reduced cell invasion obviously, moderately increased apoptosis rate, effectively inhibited the proliferation of Ishikawa cells, and siRNA-VEGF plus with Paclitaxel effected most significantly. Paclitaxel group and siRNA-VEGF plus with paclitaxel group cells were arrested in the G2/M phase, siRNA-VEGF transfected cells arrested in GO/G1phase. But these effects did not appear in scrambled siRNA(siRNA-N) control group and control group.Conclusion:The siRNA targeting humanVEGFgene could reduce VEGF expression effectively; siRNA-VEGF transfected cells in combination with Paclitaxel have synergistic effects, can significantly reduce the expression of VEGF, inhibit cellular proliferation and invasion, cells are arrested at G2phase and promote apoptosis; Paclitaxel in addition to a general anti-cancer effects but also lower the expression of VEGF, can be used as a new inhibitor of vascular.