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

M. Fujita, K. Goto, K. Yoshida, Hirohiko Okamura, H. Morimoto, S. Kito, J. Fukuda, T. Haneji

Research output: Contribution to journalArticle

29 Citations (Scopus)

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 IκB-α in whole cell lysates decreased, while the level of NF-κB in nucleus increased, in the okadaic acid-treated cells. Okadaic acid-treatment also alters the cellular localization of NF-κB, from cytoplasm to nuclei. To investigate the activation of NF-κB in okadaic acid-treated SCC-25 cells, we performed electrophoretic mobility gel shift assay using nuclear extracts and the consensus oligonucleotide for NF-κB DNA binding site. The binding of nuclear proteins to the oligonucleotide of NF-κB 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-κB 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-κB 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.

Original languageEnglish
Pages (from-to)199-206
Number of pages8
JournalOral Oncology
Volume40
Issue number2
DOIs
Publication statusPublished - Feb 2004
Externally publishedYes

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CD95 Antigens
Okadaic Acid
Fas Ligand Protein
NF-kappa B
Squamous Cell Carcinoma
Oligonucleotides
Binding Sites
Messenger RNA
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Luciferases
Reporter Genes
Genetic Promoter Regions
Carrier Proteins
Cytoplasm
Proteins
Cell Death
Western Blotting
Gels

Keywords

  • Apoptosis
  • Fas ligand
  • Fas receptor
  • NF-κB
  • Okadaic acid
  • SCC cells

ASJC Scopus subject areas

  • Oncology

Cite this

Okadaic acid stimulates expression of Fas receptor and Fas ligand by activation of nuclear factor kappa-B in human oral squamous carcinoma cells. / Fujita, M.; Goto, K.; Yoshida, K.; Okamura, Hirohiko; Morimoto, H.; Kito, S.; Fukuda, J.; Haneji, T.

In: Oral Oncology, Vol. 40, No. 2, 02.2004, p. 199-206.

Research output: Contribution to journalArticle

Fujita, M. ; Goto, K. ; Yoshida, K. ; Okamura, Hirohiko ; Morimoto, H. ; Kito, S. ; Fukuda, J. ; Haneji, T. / Okadaic acid stimulates expression of Fas receptor and Fas ligand by activation of nuclear factor kappa-B in human oral squamous carcinoma cells. In: Oral Oncology. 2004 ; Vol. 40, No. 2. pp. 199-206.
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