The Biological Effect Of Silencing SnoN Gene On Osteosarcoma MG-63 Cells And Its Primary Mechanism

Objective:Small interfering RNA(siRNA)was used to down-regulate the expression of SnoN gene,to explore the effect of silencing SnoN gene on the biological behavior of osteosarcoma MG-63 cells,and to investigate the initial mechanism of its occurrence.Methods: First,there were three groups,namely the experimental group(SnoN-siRNA group),the negative control group(control-siRNA group or NC-siRNA group)and the blank control group(non-transfected group).In the SnoN-siRNA group,liposome transfection was used to transfer the pre-designed SnoN-siRNA with silencing effect into osteosarcoma MG-63 cells.In the control-siRNA group,control siRNA(heterozygous siRNA)was transferred to osteosarcoma MG-63 cells in the same manner.No siRNA was added to the untransfected group.The proliferation of the above three groups of MG-63 osteosarcoma cells was detected by CCK-8 cytotoxic activity assay,and the migration ability of the above three groups of MG-63 cells was detected by scratch healing method.Hoechst 33258 staining was used to detect the apoptosis of osteosarcoma MG-63 cells.The expression of cyclin D1,PCNA,MMP-2,MMP-9,Bax,Bcl-2,caspase-3,caspase-9,β-catenin and c-myc in osteosarcoma MG-63 cells were examined by Western blot.Results: After transfection of SnoN-siRNA into osteosarcoma MG-63 cells,the expression level of SnoN protein in the SnoN-siRNA group was significantly down-regulated compared with that in the control-siRNA group and the non-transfected group(P<0.05).The CCK-8 test showed that OD values at 24,48 and 72 hours in the SnoN-siRNA group were significantly lower than those in the control-siRNA group and the non-transfected group(P<0.05).The results of the scratch healing experiment showed that the scratch healing rate in the SnoN-siRNA group was significantly lower than that in the control-siRNA group and the non-transfected group(P<0.05).Hoechst 33258 staining showed that the apoptosis rate of cells in the SnoN-siRNA group was significantly increased compared with the control-siRNA group and the non-transfected group(P<0.05).Western blot results showed that the relative expression levels of cyclin D1,PCNA,mmp-2,mmp-9,n-catenin,c-myc and bcl-2 in the SnoN-siRNA group were significantly lower than those in the control-siRNA group and the non-transfected group(P<0.05),while the expression levels of Bax,caspase-3 and caspase-9 in the SnoN-siRNA group were significantly lower than those in the control-siRNA group and the non-transfected group,Significantly increased(P<0.05).Conclusion:(1)After silencing the expression of SnoN gene,the proliferation,invasion and migration of osteosarcoma MG-63 cells decreased.(2)Silencing of SnoN gene expression can induce apoptosis of osteosarcoma MG-63 cells.(3)SnoN gene can regulate the above biological behavior of osteosarcoma MG-63 cells,which may be related to the Wnt/ β-catenin signaling pathway.