Activation of AMP-activated protein kinase and phosphorylation of glycogen synthase kinase3 β mediate ursolic acid induced apoptosis in HepG2 liver cancer cells

Phytother Res. 2013 Nov;27(11):1714-22. doi: 10.1002/ptr.4925. Epub 2013 Jan 16.

Abstract

Despite the antitumour effect of ursolic acid observed in several cancers, the underlying mechanism remains unclear. Thus, in the present study, the roles of AMP-activated protein kinase (AMPK) and glycogen synthase kinase 3 beta (GSK3β) were examined in ursolic acid induced apoptosis in HepG2 hepatocellular carcinoma cells. Ursolic acid significantly exerted cytotoxicity, increased the sub-G1 population and the number of ethidium homodimer and terminal deoxynucleotidyl transferase(TdT) mediated dUTP nick end labeling positive cells in HepG2 cells. Also, ursolic acid enhanced the cleavages of poly-ADP-ribose polymerase (PARP) and caspase3, attenuated the expression of astrocyte elevated gene (AEG1) and survivin in HepG2 cells. Interestingly, ursolic acid increased the phosphorylation of AMPK and coenzyme A carboxylase and also enhanced phosphorylation of GSK3β at inactive form serine 9, whereas ursolic acid attenuated the phosphorylation of AKT and mTOR in HepG2 cells. Conversely, AMPK inhibitor compound C or GSK3β inhibitor SB216763 blocked the cleavages of PARP and caspase 3 induced by ursolic acid in HepG2 cells. Furthermore, proteosomal inhibitor MG132 suppressed AMPK activation, GSK3β phosphorylation, cleaved PARP and deceased AEG-1 induced by ursolic acid in HepG2 cells. Overall, our findings suggest that ursolic acid induced apoptosis in HepG2 cells via AMPK activation and GSK3β phosphorylation as a potent chemopreventive agent.

Keywords: AMPK; GSK3β; HepG2 cells; apoptosis; ursolic acid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / metabolism*
  • Apoptosis / drug effects*
  • Carcinoma, Hepatocellular / pathology
  • Caspase 3 / metabolism
  • Cell Adhesion Molecules / metabolism
  • Glycogen Synthase Kinase 3 / metabolism*
  • Glycogen Synthase Kinase 3 beta
  • Hep G2 Cells
  • Humans
  • Indoles / pharmacology
  • Leupeptins / pharmacology
  • Liver Neoplasms / pathology
  • Maleimides / pharmacology
  • Membrane Proteins
  • Phosphorylation
  • Poly(ADP-ribose) Polymerases / metabolism
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins c-akt / metabolism
  • RNA-Binding Proteins
  • TOR Serine-Threonine Kinases / metabolism
  • Triterpenes / pharmacology*
  • Ubiquitination
  • Ursolic Acid

Substances

  • Cell Adhesion Molecules
  • Indoles
  • Leupeptins
  • MTDH protein, human
  • Maleimides
  • Membrane Proteins
  • Protein Kinase Inhibitors
  • RNA-Binding Proteins
  • SB 216763
  • Triterpenes
  • Poly(ADP-ribose) Polymerases
  • MTOR protein, human
  • GSK3B protein, human
  • Glycogen Synthase Kinase 3 beta
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Glycogen Synthase Kinase 3
  • AMP-Activated Protein Kinases
  • CASP3 protein, human
  • Caspase 3
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde