Crucial role of c-Jun phosphorylation at Ser63/73 mediated by PHLPP protein degradation in the cheliensisin a inhibition of cell transformation

Cancer Prev Res (Phila). 2014 Dec;7(12):1270-81. doi: 10.1158/1940-6207.CAPR-14-0233. Epub 2014 Oct 3.

Abstract

Cheliensisin A (Chel A), as a novel styryl-lactone isolated from Goniothalamus cheliensis Hu, has been demonstrated to have an inhibition of EGF-induced Cl41 cell transformation via stabilizing p53 protein in a Chk1-dependent manner, suggesting its chemopreventive activity in our previous studies. However, its underlying molecular mechanisms have not been fully characterized yet. In the current study, we found that Chel A treatment could increase c-Jun protein phosphorylation and activation, whereas the inhibition of c-Jun phosphorylation, by ectopic expression of a dominant-negative mutant of c-Jun, TAM67, reversed the Chel A inhibition of EGF-induced cell transformation and impaired Chel A induction of p53 protein and apoptosis. Moreover, our results indicated that Chel A treatment led to a PHLPP downregulation by promoting PHLPP protein degradation. We also found that PHLPP could interact with and bind to c-Jun protein, whereas ectopic PHLPP expression blocked c-Jun activation, p53 protein and apoptotic induction by Chel A, and further reversed the Chel A inhibition of EGF-induced cell transformation. With the findings, we have demonstrated that Chel A treatment promotes a PHLPP protein degradation, which can bind to c-Jun and mediates c-Jun phosphorylation, and further leading to p53 protein induction, apoptotic responses, subsequently resulting in cell transformation inhibition and chemopreventive activity of Chel A.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Proliferation
  • Cell Transformation, Neoplastic / drug effects*
  • Cell Transformation, Neoplastic / pathology
  • Cells, Cultured
  • Epidermal Cells
  • Epidermal Growth Factor / pharmacology
  • Epidermis / drug effects*
  • Epidermis / metabolism
  • Epoxy Compounds / pharmacology*
  • Flow Cytometry
  • Immunoprecipitation
  • Mice
  • Nuclear Proteins / metabolism*
  • Phosphoprotein Phosphatases / metabolism*
  • Phosphorylation / drug effects
  • Proteolysis / drug effects*
  • Proto-Oncogene Proteins c-jun / metabolism*
  • Pyrones / pharmacology*
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Serine / metabolism
  • Signal Transduction / drug effects*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Epoxy Compounds
  • Nuclear Proteins
  • Proto-Oncogene Proteins c-jun
  • Pyrones
  • RNA, Messenger
  • Tumor Suppressor Protein p53
  • cheliensisin A
  • Serine
  • Epidermal Growth Factor
  • PHLPP1 protein, mouse
  • Phosphoprotein Phosphatases