Activation of NFkappaB is necessary for IL-1beta-induced cyclooxygenase-2 (COX-2) expression in human gingival fibroblasts

Mol Cell Biochem. 2000 Jun;209(1-2):113-8. doi: 10.1023/a:1007155525020.


The immediate-early cyclooxygenase-2 (COX-2) gene encodes an inducible prostaglandin synthase enzyme which is implicated in inflammatory and proliferative diseases. COX-2 is highly induced during cell activation by various factors, including mitogens, hormones and cytokines. Since pro-inflammatory cytokine IL-1beta has been shown to induce prostaglandin E2 (PGE2) release in human gingival fibroblasts (HGF), here we analyzed the effect of IL-1beta on the expression of COX-2 and the activation of NFkappaB in HGF. Northern hybridization analysis revealed that IL-1beta (200 pg/ml) increased the expression of COX-2 mRNA in HGF. The effect of IL-1beta was abrogated by herbimycin A, a protein tyrosine kinase inhibitor, and enhanced by orthovanadate, a protein tyrosine phosphatase inhibitor. IL-1beta-induced PGE2 release was blocked by the tyrosine kinase inhibitor and increased by the tyrosine phosphatase inhibitor. The results of transient transfection assays using chimeric constructs of the human COX-2 promoter (nt -1432 approximately +59) ligated to a luciferase reporter gene indicated that IL-1beta stimulated the transcriptional activity approximately 1.5-fold. Gel mobility shift assays with a radiolabelled COX-2-NFkappaB oligonucleotide (nts-223 to-214) revealed an increase in the binding of nuclear proteins from IL-1beta-stimulated HGF. This increase of DNA-protein complex formation induced by IL-1beta was blocked by herbimycin A and another tyrosine kinase inhibitor, genistein. These results suggest that NFkappaB is an important transcription factor for IL-1beta-induced COX-2 gene expression, and is involved in inducing COX-2 gene transcription through tyrosine phosphorylation in HGF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cells, Cultured
  • Cyclooxygenase 2
  • Dinoprostone / metabolism
  • Fibroblasts / cytology
  • Fibroblasts / enzymology
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Expression Regulation, Enzymologic / physiology*
  • Gingiva / cytology
  • Gingiva / enzymology*
  • Humans
  • Interleukin-1 / pharmacology*
  • Isoenzymes / genetics
  • Membrane Proteins
  • NF-kappa B / metabolism*
  • Prostaglandin-Endoperoxide Synthases / genetics*
  • Protein Tyrosine Phosphatases / antagonists & inhibitors
  • RNA, Messenger / genetics
  • Rats
  • Recombinant Proteins / biosynthesis
  • Transcription, Genetic / drug effects
  • Transfection
  • Vanadates / pharmacology


  • Interleukin-1
  • Isoenzymes
  • Membrane Proteins
  • NF-kappa B
  • RNA, Messenger
  • Recombinant Proteins
  • Vanadates
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • Protein Tyrosine Phosphatases
  • Dinoprostone