AMPK activity and cell proliferation in HCC cell lines, PLC/PRF/5 and HepG2.

<p>(A): Western blot analysis of phosphor-AMPK activation induced by AICAR (1 mM) and metformin (10 mM). Representative bands from three independent experiments. (B, C): Both AICAR and metformin inhibited cell proliferation in a time- and dose-dependent manner (MTT method) in PLC/PRF/5 and HepG2. *p<0.05 (vs. control). (D): BrdU incorporation was significant decreased in both of PLC/PRF/5 and HepG2 cells by metformin treatment for 24 h (E): Metformin inhibited cell proliferation and the growth inhibition was rescued by AMPKα1 siRNA and compound C (an AMPK inhibitor) treatment (cell count method). *p<0.05 (vs. metformin only). (F): As a control, LKB1 (the upstream activator of AMPK) absent cell line, Hela cells did not show growth inhibition with AICAR and metformin treatment. (G): Effect of AMPK activators on cell cycle arrest in HCC cells. Representative DNA histograms for PLC/PRF5 cells are shown. Data from three independent experiments are shown as mean ± SD. (H): Western blot analysis of p21^CIP, p27^KIP and cyclin D1 by AICAR (1 mM) and metformin (10 mM) treatment for 24 hours in PLC/PRF/5 cells.