Effect of Nobiletin on the malignant biological behaviors of liver cancer cells by regulating the CDK1/CCNB1/PLK1 signaling pathway
Objective To investigate the effect of Nobiletin(NOB)on the malignant biological behaviors of liver cancer cells by regulating the cyclin dependent kinase 1(CDK1)/cyclin B1(CCNB1)/polo like kinase 1(PLK1)signaling pathway.Methods Human liver cancer cells(Huh-7)were treated with NOB at a concentration of 10.0-160.0μmol/L,followed by the screening of the optimal concentration by cell counting kit-8(CCK-8)assay.Huh-7 cells were treated with blank control(Control group),low-dose NOB(L group),medium-dose NOB(M group),high-dose NOB(H group),and CDK1/CCNB1 inhibitor(CE group).Cell proliferation,apoptosis,migration,and invasion were examined by colony formation assay,flow cytometry,wound healing assay and Transwell assay respectively.Western blot was applied to detect the protein expressions of proliferation-related proteins(Ki67,Cyclin D1),apoptosis-related proteins(Bax,Caspase-3),migration/invasion-related proteins(MMP-2,MMP-9),CDK1,CCNB1,and PLK1.A nude mouse xenograft model was created to detect the in vivo effect of NOB on the growth of liver cancer.Results NOB concentrations of 20pmol/L,40μmol/L,and 80μmol/L were selected as the low-dose,medium-dose and high-dose treatment for subsequent experiments,respectively.Compared with those of the Control group,cells in the L,M,H,and CE groups showed significantly lower number of colonies,wound healing rate,invasive cell number,protein levels of Ki67,Cyclin D1,MMP-2,MMP-9,CDK1,CCNB1,and PLK1,and higher apoptotic rate and protein levels of Caspase-3,and Bax(P＜0.05).In the nude mouse xenograft model,NOB treatment yielded a significantly slower tumor growth,lower tumor weight and volume and lower protein levels of CDK1,CCNB1,and PLK1(P＜0.05).Conclusion NOB inhibits the malignant biological behavior of liver cancer cells by inactivating the CDK1/CCNB1/PLK1 signaling pathway.
万方数据
维普
