| Objective: To investigate the relationship between Wnt/?-catenin/TCF-4 pathway and livin and the mechanisms of apoptotic regulation in renal cell carcinoma by inhibiting Wnt pathway with exogenous indomethacin.Methods: Renal cancer cell line 786-0 was used in the study.786-0cells were treated with different concentrations of indomethacin.The proliferation activity of cells was tested by CCK8 method.Flow cytometry analysis were used to determine cell apoptosis.The expressions of mRNA of ?-catenin,TCF-4,caspase-3 and livin were detected by real-time PCR.The protein expression levels of ?-catenin,caspase-3 and livin were assessed by Western blot.Results: After indomethacin intervention,the cell viability decreased significantly in a dose-effect relationship(P=0.000).Flow pattern showed that with the increase of indomethacin concentration,the apoptotic rate increased gradually.The apoptotic rates of 25?mol/L group,50?mol/Lgroup,100?mol/L group were(32.07+1.01)%,(40.03+1.95)% and(72.33+0.02)% respectively which have significant differences compared to the control group(P=0.000,P=0.002,P=0.000).Real-time PCR results showed that the relative expression levels of ?-catenin,TCF-4 and livin decreased 24 hours after indomethacin intervention(P=0.002,P=0.000,P=0.000),while the relative expression of caspase-3 increased significantly(P=0.001).Western blot analysis showed that in the 25?mol/L group,50?mol/L group,100?mol/L group,the relative expression levels of?-catenin were 0.568±0.020(P=0.001),0.396±0.030(P=0.000),0.142±0.038(P=0.000);the relative expression of livin were 0.139±0.016(P=0.005),0.050±0.006(P=0.000),0.011±0.0019P=0.000);and the relative expression of caspase-3 were 0.278±0.035(P=0.046),0.396±0.071(P=0.000),0.518±0.015(P=0.000).Conclusion: Degradation of ?-catenin/TCF-4 may lead to the transcriptional inhibition of livin and upregulation of caspase-3,and then induces apoptosis of renal cancer cells. |
PDF Full Text Request