Okadaic acid stimulates expression of Fas receptor and Fas ligand by activation of nuclear factor kappa-B in human oral squamous carcinoma cells

Oral Oncol. 2004 Feb;40(2):199-206. doi: 10.1016/s1368-8375(03)00152-0.

Abstract

In the present study, we used western blot and RT-PCR analysis to examine the expression of proteins and mRNAs of Fas receptor and Fas ligand in human oral squamous carcinoma SCC-25 cells treated with okadaic acid. Treatment with okadaic acid enhanced the expression of proteins and mRNAs of both Fas receptor and Fas ligand in SCC-25 cells. The amount of IkappaB-alpha in whole cell lysates decreased, while the level of NF-kappaB in nucleus increased, in the okadaic acid-treated cells. Okadaic acid-treatment also alters the cellular localization of NF-kappaB, from cytoplasm to nuclei. To investigate the activation of NF-kappaB in okadaic acid-treated SCC-25 cells, we performed electrophoretic mobility gel shift assay using nuclear extracts and the consensus oligonucleotide for NF-kappaB DNA binding site. The binding of nuclear proteins to the oligonucleotide of NF-kappaB increased when the cells had been treated with 20 nM okadaic acid for 4 h. We transfected the cells with pFLF1, which has the promoter region of Fas receptor gene containing NF-kappaB binding site. A luciferase reporter gene assay demonstrated that the activity in the cells transfected with pFLF1 and treated with 20 nM okadaic acid increased in a time-dependent manner and that the activity was more than three-fold over that in the control cells. Our results suggest that NF-kappaB activated at early stages in the okadaic acid-treated SCC-25 cells stimulated the promoter activity of Fas receptor in the cells leading to the apoptotic death of these cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Squamous Cell / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Fas Ligand Protein
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • I-kappa B Proteins / metabolism
  • Ligands
  • Membrane Glycoproteins / drug effects*
  • Membrane Glycoproteins / metabolism
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / drug effects
  • NF-kappa B / metabolism
  • Neoplasm Proteins / metabolism
  • Okadaic Acid / pharmacology*
  • Promoter Regions, Genetic
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tongue Neoplasms / metabolism*
  • Transfection
  • Tumor Cells, Cultured
  • fas Receptor / drug effects*
  • fas Receptor / genetics
  • fas Receptor / metabolism

Substances

  • Enzyme Inhibitors
  • FASLG protein, human
  • Fas Ligand Protein
  • I-kappa B Proteins
  • Ligands
  • Membrane Glycoproteins
  • NF-kappa B
  • NFKBIA protein, human
  • Neoplasm Proteins
  • fas Receptor
  • NF-KappaB Inhibitor alpha
  • Okadaic Acid