The Effect Of RNAi Inhibition Of GSK3? Expression On The Growth Of Human Glioma Cells And Its Mechanism

[Object] glioma is one of the most fatal tumors,so the urgent affairs is to take advantage of the new molecular targets to improve the current treatment means.In this experiment,we first construct siRNA expression vector for the targeting,which lays a good foundation for the follow-up study on the role of GSK3? in the genesis and development of glioma,and observe its affect to the proliferation,apoptosis,invasion ability of the human brain glioma cell lines U87 after inhibiting the expression of GSK3?.[Method] Designing the GSK3? siRNA sequence following the design philosophy for siRNA,synthesizing its expression frame and connect it with the vector p FU-GW-RNAi,constructing the overexpression vector for GSK3? at the same time,at last,cotransfecting the 293T-cell,detecting the GSK3? protein expression in the 293 T cell after transfecting by means of the western-blot,to screen out the target spot sequeen with the highest inhibition ration.Introducting the targeting siRNA expression vector for the GSK3? gene into the U87 cellin vitro,detecting the inhibiton ratio by means of the western-blot and detecting the change of the biological characteristic by means of the transwell experiment?MTT experiment?Flow cytometry.[Result] The sequencing data demonstrated the successful construct of the recombinant plasmid,the No.4 target sequence with the most inhibition ratio to the GSK3? gene was sceened,a targeting siRNA expression vector for the GSK3? gene was constructed.Infecting the U87 cell with the targeting GSK3? siRNA construct of which the vector may downregulate the expression of the GSK3? gene,the invasion ability,the multiplication capacity of the U87 cell obviously,and increase the apoptosis ratio of the U87 cell.[Conclusion] The constructed siRNA expression vector for the targeting GSK3? gene may inhibit the expression of the GSK3? gene effectively,and the inhibition to the GSK3? gene in the U87 cell of the human glioma cell line may inhibit the multiplication and the invasion abilities prominently and increase the apoptosis ratio of the U87 cell.It is demonstrated that GSK3? gene may play a very important role during the genesis and development of the glioma,which provide a new research idea for the gene therapy for the glioma.