Short Hairpin Rna Inhibits The Expression Of Livin Gene In Mcf7 Cells

Objective: To explore livinα-specific shRNA inhibiting the express of livinαinMCF7 cells.Methods: To construct four groups vector(Pgenesil-8-livin-shRNA1, Pgenesil-8-livin-shRNA2, Pgenesil-8-livin-HK, Pgenesil-8-livin), which express EGFP-Livin.(Pgenesil-8 isa plamid product; shRNA1, shRNA2 are two RNAi sequences; livin stands for the cDNA oflivina GI: 15680240; HK is a negative sequence)The four groups vector transfected intoMCF7 cells. After 24h,to search green fluorescent.After 48h,to take photos for four groupscell,and detect the expression of EGFP by FACS,and then to detect the mRNA of livinαby RT-PCR.Results: The four vectors transfected into cells succedly, and 40%-70% cells weretransfected. The cells'fluorescent of the two groups cell,into which were transfectedPgenesil-8-livin-shRNA1, Pgenesil-8-livin-shRNA2 are lighter than another two groups.Thelevel of mRNA in the group of Pgenesil-8-livin-shRNA2 is lower than the group ofPgenesil-8-livin, and the difference is significant(P＜0.01). But the group ofPgenesil-8-livin-shRNA1 compare with Pgenesil-8-livin the mRNA level has littlediffenence(P＞0.05).conclusions: The interference sequence of shRNA2 is effect to livinαin MCF7cells,and it can inhibit the expression of livinα, but shRNA1 can't, shRNA2 can interfere theexpression of livinαoutside of MCF7 cell. Base on it we can do interference experimentinside cell.