Extracellular NAMPT/visfatin causes p53 deacetylation via NAD production and SIRT1 activation in breast cancer cells

Cell Biochem Funct. 2017 Aug;35(6):327-333. doi: 10.1002/cbf.3279.

Abstract

Visfatin, which is secreted as an adipokine and cytokine, has been implicated in cancer development and progression. In this study, we investigated the NAD-producing ability of visfatin and its relationship with SIRT1 (silent information regulator 2) and p53 to clarify the role of visfatin in breast cancer. MCF-7 breast cancer cells were cultured and treated with visfatin. SIRT1 activity was assessed by measuring fluorescence intensity from fluoro-substrate peptide. To investigate the effect of visfatin on p53 acetylation, SDS-PAGE followed by western blotting was performed using specific antibodies against p53 and its acetylated form. Total NAD was measured both in cell lysate and the extracellular medium by colorimetric method. Visfatin increased both extracellular and intracellular NAD concentrations. It also induced proliferation of breast cancer cells, an effect that was abolished by inhibition of its enzymatic activity. Visfatin significantly increased SIRT1 activity, accompanied by induction of p53 deacetylation. In conclusion, the results show that extracellular visfatin produces NAD that causes upregulation of SIRT1 activity and p53 deacetylation. These findings explain the relationship between visfatin and breast cancer progression.

Keywords: SIRT1; acetylation; breast cancer; p53; visfatin.

MeSH terms

  • Acrylamides / pharmacology
  • Breast Neoplasms
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Humans
  • MCF-7 Cells
  • NAD / metabolism*
  • Nicotinamide Phosphoribosyltransferase / genetics
  • Nicotinamide Phosphoribosyltransferase / metabolism
  • Nicotinamide Phosphoribosyltransferase / pharmacology*
  • Piperidines / pharmacology
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / pharmacology
  • Sirtuin 1 / metabolism*
  • Tumor Suppressor Protein p53 / metabolism*
  • Up-Regulation / drug effects*

Substances

  • Acrylamides
  • N-(4-(1-benzoylpiperidin-4-yl)butyl)-3-(pyridin-3-yl)acrylamide
  • Piperidines
  • Recombinant Proteins
  • Tumor Suppressor Protein p53
  • NAD
  • Nicotinamide Phosphoribosyltransferase
  • SIRT1 protein, human
  • Sirtuin 1