Gut 50:771-778 doi:10.1136/gut.50.6.771
  • Helicobacter pylori

Analysis of apoptotic and antiapoptotic signalling pathways induced by Helicobacter pylori

  1. S Maeda,
  2. H Yoshida,
  3. Y Mitsuno,
  4. Y Hirata,
  5. K Ogura,
  6. Y Shiratori,
  7. M Omata
  1. Department of Gastroenterology, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan
  1. Correspondence to:
    S Maeda, Department of Gastroenterology, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo113-8655, Japan;
  • Accepted 18 September 2001


Background and aims: Although it is reported that Helicobacter pylori induces apoptosis on gastric epithelial cells, the mechanism remains unknown. Antiapoptotic effects generated by H pylori have not yet been evaluated.

Methods: (1) H pylori strains (type 1 wild, TN2-ΔcagE, TN2-ΔvacA) were cocultured with MKN45, TMK1, and HeLa cells, and cell viability and apoptosis were assessed by trypan blue exclusion and DNA laddering, respectively. (2) Activation of caspases-3, 7, and 8, cytochrome c release from the mitochondria, and Fas, Fas associated death domain protein (FADD), Bax, Bak, and Bcl-X expression were evaluated by immunoblot analysis. (3) To investigate whether nuclear factor kappa B (NFκB) activation induced by cag pathogenicity island (PAI) positive H pylori affects antiapoptosis, MKN45 cells stably expressing super-repressor Iκβα were cocultured with H pylori, and cell viability and caspase activation were evaluated. NFκB regulated gene expression was also evaluated by ribonuclease protection assay.

Results: (1) Wild-type and ΔvacA mutant H pylori induced apoptosis more potently than the ΔcagE mutant. Inhibition of cell contact between H pylori and cancer cells and heat killing H pylori diminished cell death. (2) Caspases-3, 7, and 8 were activated time dependently by H pylori as well as by the agonist anti-Fas. Cytochrome c release from mitochondria was observed and was not inhibited by caspase-8 inhibitor. Although protein expression of Fas, FADD, Bax, Bak, and Bcl-X in the whole cell lysates was not changed by H pylori, Bax was decreased from mitochondria free cytosol suggesting that Bax was translocated into mitochondria. (3) Cell death and the activities of caspases-3 and 8 were promoted in MKN45 cells stably expressing super-repressor Iκβα that inhibits NFκB activation. Antiapoptotic proteins c-IAP1 and c-IAP2 were upregulated by the wild-type strains.

Conclusion:cag PAI positive H pylori is capable of inducing apoptotic effects mainly through the mitochondrial pathway. Antiapoptotic effects mediated by NFκB activation were also observed.