Study Of The Effects Of Ribozyme And SiRNA Targeting Survivin In Inducing Apoptosis Of Tumor Cells

Objective: To investigate the gene silencing and apoptosis-inducing effects of pRNA/Ribozyme(survivin) and pRNA/siRNA(survivin) chimera.Methods: Construct pRNA/Ribozyme(survivin) and pRNA/siRNA(survivin) chimera. Lipotransfect the chimera into human cancer cell line: MCF-7 and Hela. Examine the mRNA level of survivin within the cancer cell line by real-time PCR. Monitor the apoptosis-inducing effects of the chimera with inverse microscope and flow cytometry after Annexin-V/PI double-labeling.Results: Investigation with celluar model in vitro revealed that:â‘ After transfection of pRNA/Ribozyme(survivin),the mRNA level of survivin within human breast cancer cell line MCF-7 was significantly inhibited compared to pRNA/Ribozyme(mutant) transfected group,and no distinctively difference in mRNA level was found between the mutant chimera transfected cells and the non-transfected.â‘¡The apoptosis rate is higher in pRNA/RZ(survivin) and pRNA/siRNA(survivin) treated cells compared to control.â‘¢After con-transfection of GFP expression plasmid and pRNA/siRNA(GFP), pRNA/siRNA(GFP)was proved to be able to significantly inhibit GFP expression by using inverse fluorescence microscope and flow cytometry,while pRNA/siRNA(survivin) having no silencing effects on GFP expression which showed that pRNA/siRNA(survivin) chimera interfered gene expression specifically.Conclusions: pRNA/Ribozyme(survivin) and pRNA/siRNA(survivin) chimera was testified to be able to silence survivin expression and induce apoptosis of cancer cell lines specifically which proved that pRNA could be used as effective vectors for small gene therapy molecules.