In Vitro Study Of RNA Interference On STAT6 Gene

Asthma is associated with airway hyperreactivity and allergic peribronchial inflammation. The immune pathogenesis of asthma is Th1/Th2 cell disfunction. STAT6 play a important role in Th2 classifacation. STAT6 is transcription factor in Th2 differentiation,which can increase Th2 cytokines such as IL-4,IL-13. STAT6 specifical induce the shift from Th cell to Th2 cell. There is no report the siRNA sequene of STAT6. So we take STAT6 gene as RNA interference target. By constructing STAT6 interference plasmid , to provide a new idea for gene therpy on asthma.Objective: To construct the recombinant plasmid expressing STAT6 shRNA and observe the effects of RNAi mediated STAT6 gene silencing.Methods: shRNA , synthesized by transcription in vitro , and the plasmid expressing STAT6 gene and EGFP were cotransfected into COS7 cell , to express the STAT6 protein and investigate the suppression effect of shRNA on STAT6 under fluorescent microscope . This effect was confirmed by western blot at expression level.Results: STAT6 protein exhibitited hign expression in COS7 cells after transfecting pIRES2– EGFP– STAT6 plasmid into COS7 cells along ; compared with the normal control group, the inhibitory rate in the two group were(74.3±5.9%)and(85.8±6.3%). pGenesil3-STAT62 was more efficacious in suppressing STAT6 protein expression.Conclusion: STAT6 shRNA were successfully constructed. STAT6 may be a promising target of gene therpy on asthma.