| Objective:To investigate the effects of Wnt/?-catenin signaling pathway inhibitor IWR-1 on proliferation,migration of human osteosarcoma cell lines MG-63,and to explore the possible mechanism.Methods:MG-63 cells were treated with IWR-1(2,4,8,16?mol/L)of different concentration gradient.CCK-8 assay was used to detect the growth inhibition rates of MG-63 cells.Cell cycle and apoptosis rate of MG-63 cells were detected by flow cytometry(FCM).The migration ability of MG-63 cells was abserved by wound scratch assay.The expressions of ?-catenin and Cyclin D1 mRNA in MG-63 cells were respectively detected by real time polymerase chain reaction(RT-PCR)and Western blot.Expression of Axin 1 and p-?-catenin in MG-63 cells was detected by Western blot.Results:Compared with the control group without IWR-1,CCK-8 assay showed the proliferation of MG-63 cells was significantly inhibited treated with IWR-1(P<0.05),and the growth inhibition of IWR-1 on MG-63 cells changed with dose and time(P<0.05).According to the results of FCM,the cell cycle of MG-63 cells showed significantly blocking in G0/G1(P<0.05).,and the apoptosis rate of MG-63 cells increased significantly with the changing of drug concentration(P<0.01),with a dose-dependent effect.IWR-1 could slow down the scratch healing of MG-63 cells,MG-63 cells treated with IWR-1 had signnificantly larger scratch area(P<0.01)and lower scratch healing rate(P<0.01)compared with control group,with a dose-dependent effect.Real-time fluorescence quantitative PCRresults showed that IWR-1 could decrease mRNA expression of ?-catenin and Cyclin D1(P<0.01).Western blot results showed that IWR-1 could increase Axin 1 and p-?-catenin protein expression(P<0.01),while ?-catenin and Cyclin D1 protein expressionwas decreased(P<0.01),showing a significant dose-dependet.Conclusion:IWR-1 can inhibit the proliferation and migration of human osteosarcoma MG-63 cells,which may be achieved by inhibiting the Wnt/?-catenin signaling pathway. |
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