RNAi-silenced Survivin and hTERT gene inhibits proliferation and induces apoptosis of human colorectal carcinoma cell line SW 480 / 基础医学与临床

Objective To investigate the Influence of Survivin and hTERT gene on cell proliferation and apoptosis in human colorectal carcinoma cell line SW 480 and to find experiment evidence for gene therapy of colorectal carci-noma .Methods Plasmids carrying shRNAs targeting survivin and hTERT were designed , constructed and trans-fected into SW480 cells.SW480 cells were then divided into blank group , blank Plasmid control group , survivin RNAi group , hTERT RNAi group and Survivin-hTERT RNAi group .The telomerase activity was examined by TRAP-PCR-ELISA analysis 48h after hTERT-shRNA transfection.Survivin and hTERT mRNA and protein expres-sion was analyzed by RT-PCR and Western blot .Cell apoptosis , proliferation were measured by flow cytometry , CCK-8 assay.Results Telomerase activity of SW480 cells in Survivin-hTERT RNAi groups were significantly decreased compared with the blank group ( P<0.01 ) .The expression of survivin and hTERT mRNA, proteins in the Survivin-hTERT RNAi group was reduced by 82.8%and 73.6%( P<0.01 ) ,79.2%and 66.7%( P<0.01 ) respectively .The inhibitory rate of cell proliferation of Survivin-hTERT RNAi group was 43.6% ±0.1%( P <0.01 ) .The apoptosis rate was 39.2%±2.3%( P<0.01 ) in the Survivin-hTERT RNAi group .Conclusions The Survivin-hTERT RNAi group could significantly reduces the protein expression of survivin and hTERT mRNA, in-hibit cell proliferation and induces cell apoptosis in human colorectal carcinoma cell line SW 480 .

Influences of shRNA on survivin gene expression and sensitivity of A549 cells to paclitaxel / 中国药理学通报

Aim To construct the eukaryotic expression vectors of short hairpin RNA targeting survivin and observe its effect on biologic behavior of A549 cells and sensitivity of A549 cells to paclitaxel.Methods The DNA fragment targeting human survivin was inserted into the plasmid,and the recombinant plasmid was constructed.The recombinant plasmids cells were transfected into A549 cells by FuGENE transfection reagent.The expression levels of survivin gene were detected with RT-PCR and Western blot before and after transfection,respectively.Cell apoptosis was detected by TUNEL method.The sensitivity of A549 cells to paclitaxel was detected by MTT after transfection.Results The recombinant plasmid was successfully constructed.RNAi group cells showed lower expression of survivin than control group.The apoptosis rate of A549 cells increased after transfection.The IC50 of paclitaxel inhibiting A549 cells was 11.9 fold before transfection compared with those after transfection.There was significant difference between the two groups(P