H. pylori activates NF-kappaB through a signaling pathway involving IkappaB kinases, NF-kappaB-inducing kinase, TRAF2, and TRAF6 in gastric cancer cells

Gastroenterology. 2000 Jul;119(1):97-108. doi: 10.1053/gast.2000.8540.

Abstract

Background & aims: H. pylori infection on gastric epithelial cells has been shown to induce NF-kappaB activation, but the mechanism of intracellular signal conduction that leads to NF-kappaB activation is not clear. The aim of this study was to analyze the molecular mechanism responsible for H. pylori-mediated NF-kappaB activation on gastric cancer cells.

Methods: NF-kappaB activation by H. pylori was tested by using luciferase reporter assay. IkappaBalpha degradation by H. pylori infection was assessed by immunoblotting. IKKalpha and IKKbeta activation was analyzed by kinase assay. In transfection experiments, effects of dominant negative IkappaBalpha, IKKalpha, IKKbeta, NF-kappaB-inducing kinase (NIK), TRAF2, and TRAF6 mutants were investigated. The effects of an IKKbeta-specific inhibitor, aspirin, on NF-kappaB activation and IL-8 secretion were also analyzed.

Results: H. pylori promotes degradation of IkappaBalpha, a cytoplasmic inhibitor of NF-kappaB. In kinase assay, H. pylori induced IKKalpha and IKKbeta catalytic activity in gastric cancer cells. Transfection of kinase-deficient mutant of either IKK inhibited H. pylori-mediated NF-kappaB activation dose-dependently. Aspirin inhibited both NF-kappaB activation and IL-8 secretion induced by H. pylori. NF-kappaB activation was also inhibited by transfection of kinase-deficient NIK or a dominant negative mutant of upstream adapter protein TRAF2 or TRAF6.

Conclusions: H. pylori induces NF-kappaB activation through an intracellular signaling pathway that involves IKKalpha, IKKbeta, NIK, TRAF2, and TRAF6.

MeSH terms

  • Aspirin / pharmacology
  • Genes, Dominant
  • Helicobacter pylori / physiology*
  • Humans
  • I-kappa B Kinase
  • I-kappa B Proteins / metabolism
  • I-kappa B Proteins / physiology
  • Interleukin-8 / metabolism
  • Mutation / physiology
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / physiology*
  • Phosphorylation
  • Phosphotransferases / deficiency
  • Protein-Serine-Threonine Kinases / physiology
  • Proteins / genetics
  • Proteins / physiology
  • Signal Transduction / physiology*
  • Stomach Neoplasms / microbiology
  • Stomach Neoplasms / pathology
  • Stomach Neoplasms / physiopathology*
  • TNF Receptor-Associated Factor 2
  • TNF Receptor-Associated Factor 6
  • Tumor Cells, Cultured

Substances

  • I-kappa B Proteins
  • Interleukin-8
  • NF-kappa B
  • Proteins
  • TNF Receptor-Associated Factor 2
  • TNF Receptor-Associated Factor 6
  • Phosphotransferases
  • Protein-Serine-Threonine Kinases
  • CHUK protein, human
  • I-kappa B Kinase
  • IKBKB protein, human
  • IKBKE protein, human
  • NF-kappa B kinase
  • Aspirin