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Casticin, a polymethoxyflavone, has been reported to exert anticancer activities.The objectives of this study were to examine the molecular mechanisms by whichcasticin induces the growth inhibition and cell cycle arrest in human hepatocellularcarcinoma (HCC) cells. The HCC cell lines Hep G2 and PLC/PRF/5 were cultured in vitro.The growth inhibitory effects of casticin were evaluated using clonogenic assays.The distribution of phases in the cell cycle was analyzed using flow cytometry(FCM) analysis with propidium iodide (PI) staining. Multiple molecular techniques,such as western blotting and gene transfection, were used to explore the molecularmechanisms of action. Our data demonstrated that casticin significantly inhibitedcell viability and colony formation in HCC cells. Furthermore, it induced cellcycle arrest in the G2/M phase. Casticin inhibited phosphorylation of the FOXO3aprotein and decreased the expression of FoxM1 and its downstream genes, such ascyclin-dependent kinase (CDK1), cdc25B and cyclin B and increased the expressionof p27KIP1. Silencing of FOXO3a expression by small interfering RNA (siRNA) transfectionclearly attenuated the inhibitory effects of casticin on FOXM1 expression andcell growth. Our findings provided clear evidence that casticin induces growthsuppression and cell cycle arrest through inhibition of FOXO3a phosphorylationcausing inactivation of FOXM1 in HCC cells.

Casticin induces growth suppression and cell cycle arrest through activation of FOXO3a in hepatocellular carcinoma