We recently observed that Compound C (CompC), a reversible inhibitor of AMP-activated protein kinase, reduced the cell viability of B16-F1 melanoma cells. To establish its molecular mechanism(s) of action, the cell cycle was examined by flow cytometry and the expression of cell cycle regulatory proteins and angiogenesis-related proteins were examined by western blot analysis. In addition, its effect on tumor growth was investigated using C57BL/6 syngeneic mice bearing B16-F1 xenografts. We found that CompC induced G2/M cell cycle arrest, which was associated with reduced levels of cell cycle regulatory proteins, such as phosphorylated pRB, cyclin-dependent protein kinases (Cdks), cyclins, and phosphorylated P-Ser10-histone H3, and increased levels of Cdk inhibitors, such as p21 and p53. We also found that CompC inhibits proliferation, migration, and tube formation of human umbilical vascular endothelial cells via the inhibition of vascular endothelial growth factor receptor-induced signaling pathways. As expected, CompC significantly reduced the tumor size of B16-F1 xenografts in the syngeneic mouse model. Inhibition of tumor growth may be attributed to reduced cell proliferation via cell cycle inhibition and in part to decreased angiogenesis in CompC-treated mice. These findings suggest the potential use of CompC against melanoma development and progression.
Keywords: B16-F1; C57BL/6; HUVECs; VEGFR; angiogenesis; cell cycle arrest; compound C.