RNA Interference Inhibited Livin Expression And Cell Proliferation Of Colon Carcinoma

Objective:To inhibit Livin expression by RNA interference in human colon carcinoma cells,and observe the effective of cell proliferation when Livin was inhibited ,so that RNA interference could be used in clinical therapy.Methods: sequences of Livin gene was retrieved from gene bank,shRNA o1igonuc1eotides were synthesized and inserted into Pgenesil-1. After DH5a strains were transformed with the vectors,plasmids were extracted,and the recombinant sequences were identified by enzyme digestion and the sequence analysis.The eukaryotic expression vectors were transfected into colon carcinoma cells(HT-29),observe green fluorescin by Fluorescence microscope. The changes of Livin expressions were detected by Western-blot. The effect of suppressing proliferation of HT-29 cell was detected by MTT assay. Cell cycles were determined by flow cytometry.Results: Three specific siRNA eukaryotic expression vectors targeting Livin were successfully constructed, which could inhibit the expression of Livin protein to varying degree,and si-Livin2 resulted in the highest inhibiting rate. The expressions of Livin protein inhibiting rates were 67.15%.Forty-eight hours after si-Livin2 transfection, the proliferation of HT-29 cells was strongly inhibited, with the inhibiting rate of 28.6%. The cell percentage of the G0/Gl phase(64.70±2.60) % was increased and the S phase(28.77±2.70) % was decreased.Conclusion:The expression of Livin could be inhibited by RNA interference . Livin-targeted RNAi can inhibit proliferation in HT-29 cells.