Transcription factor AP1 binds the functional region of the promoter and regulates gene expression of human PPARdelta in LoVo cell

Tumour Biol. 2013 Dec;34(6):3619-25. doi: 10.1007/s13277-013-0943-4. Epub 2013 Jul 6.

Abstract

Peroxisome proliferator-activated receptor δ gene (PPARδ) is correlated with carcinogenesis of colorectal cancer, but the regulation of its gene transcription remains unclear. We herein report that AP1 binds the promoter and regulates PPARδ gene expression. With a luciferase reporter system, we identified a functional promoter region of 30 bp of PPARδ gene by deletion and electrophoretic mobility shift assays (EMSA). Using site-directed mutagenesis and decoy analyses, we demonstrated that AP1 bound the functional transcriptional factor binding site in a region extending from -176 to -73 of the PPARδ promoter, which was confirmed using EMSA and supershift assays. Consequently, inhibition of the AP1 binding site led to decreased PPARδ mRNA. Our study demonstrated that AP1 is the transcriptional factor that contributes to PPARδ expression in LoVo cells.

MeSH terms

  • Anthracenes / pharmacology
  • Base Sequence
  • Binding Sites / genetics
  • Cell Line, Tumor
  • Cell Nucleus / genetics
  • Cell Nucleus / metabolism
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / metabolism
  • Colonic Neoplasms / pathology
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Regulation*
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Mutagenesis, Site-Directed
  • PPAR delta / genetics*
  • PPAR delta / metabolism
  • Promoter Regions, Genetic / genetics*
  • Protein Binding / drug effects
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor AP-1 / antagonists & inhibitors
  • Transcription Factor AP-1 / metabolism*
  • Transfection

Substances

  • Anthracenes
  • PPAR delta
  • Transcription Factor AP-1
  • pyrazolanthrone
  • Luciferases