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《Journal of Harbin Medical University》 2013-01
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Effects of siRNA targeted against Fbxw8 on cell proliferation of prostate cancer cells

ZHAO Bin-jia-hui1,LIN Ping1,CHEN Yuan-yuan1,XIAO Li-jie1,LIN Feng1,ZHENG Xiu-lan2,ZOU Hai-feng1,YU Xiao-guang1(1.Department of Biochemistry and Molecular Biology,Harbin Medical University;2.Department of Diagnosis,The Third Affiliated Hospital of Harbin Medical University,Harbin 150081,China)  
Objective To investigate the effect of F-box and WD repeat domain containing 8(Fbxw8) on cell proliferation and the expression levels of proliferation-related genes such as cyclinA,cyclinB,cdk1,cdk2,p21 and p27 in human prostate cancer DU145 cells.Methods RT-PCR and Western blot were applied to evaluate the interference efficiency of Fbxw8 siRNAs.The cell proliferation was assessed by CCK-8 assay.The cell cycle of DU-145 cells was determined by flow cytometry.The expression levels of CDK1,CDK2,cyclin A,cyclinB,P21 and P27 proteins were detected by Western blot.Results Fbxw8 siRNA significantly decreased the mRNA and protein of Fbxw8 in DU145 cells(P0.01).The proliferation of DU-145 cells was obviously inhibited after Fbxw8 siRNA intervention,especially in 72 h(P0.05).Flow cytometry analysis showed that Fbxw8 siRNA caused cell cycle arrest in G2/M phase and cell percent of G2/M phase increased from 15.32%(control siRNA) to 29.03%(Fbxw8 siRNA 1) and 28.39%(Fbxw8 siRNA 2).Westert blot showed that the levels of CDK1,CDK2,CyclinA and CyclinB expression were down-regulated whereas the levels of p21 and p27 were up-regulated.Conclusion Targeted inhibition of Fbxw8 in DU145 cells can significantly inhibit cell proliferation and block cells in G2 / M phase,which may be achieved by decreasing the expressions of CDK1,CDK2,cyclin A and cyclin B1 and increasing the expressions of P21 and P27.
【Fund】: 国家自然科学基金资助项目(81172417)
【CateGory Index】: R737.25
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