Inhibition Of GSK-3β Gene Expression By RNA Interference And Its Effect On The Nogo-A Inhibition To Axon Outgrowth Of PC12 Cells

Objective: To study the effect of GSK-3βgene silence on the Nogo-A inhibition to axon outgrowth of PC12 cells.Methods: The small hairpin RNA eukaryotic expression vector targeting GSK-3βwas constructed and cultured PC12 cells were transfected by lipofection using Lipofectamine reagent. 24 hours following tansfection, cells were examined with fluorescence microscope to observe the efficiency of transfection,the mRNA expression of GSK-3βwere evaluated by semi-quantitative reverse transcription(PCR ). And cells were treated with NGF at a concentration of 0.5mg/L to induce differentiation. 72h later, density of cells was modulated and Nogo–A was added to the cells at a concentration of 2.5mg/L. 48h later the effect of siGSK-3βon axon outgrowth in the presence of Nogo-A and axon length were measured by Image-Pro Plus6.0 software .Results: (1)The siRNA expression vector targeting GSK-3βwas constructed successfully ; (2)Compared with untransfectedcells,transfected cells emited green fluorescence transfection efficiency was 43%,80%,76% respectively in the test group,control group and negative control group ;(3)The GSK-3βgene expression was inhibited efficiently;(4) 72h following the induction of NGF,in presence of Nogo-A,treatment of PC12 cells with siGSK-3βresulted in a promotion of axon outgrowth and the axon length had a significant difference between the test group and control group . Conclusion: GSK-3βplays a role in the Nogo-A inhibition on axon outgrowth of PC12 cell.