Helicobacter pylori and H2O2 increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells

Song Ze Ding, Ann M. O'Hara, Tim L. Denning, Bernadette Dirden-Kramer, Randy C. Mifflin, Victor E. Reyes, Kieran A. Ryan, Susan N. Elliott, Tadahide Izumi, Istvan Boldogh, Sankar Mitra, Peter B. Ernst, Sheila E. Crowe

Research output: Contribution to journalArticle

71 Scopus citations

Abstract

Background & Aims: Helicobacter pylori infection causes inflammation, accumulation of reactive oxygen species, and oxidative DNA damage in the gastric mucosa. Apurinic/apyrimidinic endonuclease-1 (APE-1)/redox factor-1 (Ref-1) repairs damaged DNA and reductively activates transcription factors, including activator protein-1. Considering that H. pylori generate reactive oxygen species and that reactive oxygen species modulate APE-1/Ref-1 in other cell types, we examined the effect of H. pylori, oxidative stress, and antioxidants on APE-1/Ref-1 expression in human gastric epithelial cells. Methods: Human gastric epithelial cell lines or cells isolated from mucosal biopsy samples were stimulated with H. pylori, Campylobacter jejuni, and/or H 2O 2 in the presence or absence of antioxidants. APE-1/Ref-1 expression was assayed by Western blot or reverse-transcription polymerase chain reaction, and its cellular distribution was determined by using indirect conventional and confocal immunofluorescence. New protein synthesis was detected by [S 35]methionine labeling. APE-1/Ref-1 function was assessed by using a luciferase-linked reporter construct containing 3 activator protein 1 binding sites. Results: APE-1/Ref-1 protein and messenger RNA were detected in resting gastric epithelial cells. APE-1/Ref-1 protein expression was increased after stimulation with H 2O 2 or live cag pathogenicity island-bearing H. pylori, but not cag pathogenicity island-negative H. pylori or C. jejuni. H. pylori- or reactive oxygen species-mediated increases in APE-1/Ref-1 expression involved de novo protein synthesis that was inhibited by antioxidants. H. pylori or H 2O 2 also induced nuclear accumulation of APE-1/Ref-1, and overexpression of APE-1/Ref-1 increased activator protein 1 binding activity. Conclusions: The data show that H. pylori or reactive oxygen species enhance APE-1/Ref-1 protein synthesis and nuclear accumulation in human gastric epithelial cells and implicate APE-1/Ref-1 in the modulation of the pathogenesis of H. pylori infection.

Original languageEnglish (US)
Pages (from-to)845-858
Number of pages14
JournalGastroenterology
Volume127
Issue number3
DOIs
StatePublished - Sep 2004

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

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    Ding, S. Z., O'Hara, A. M., Denning, T. L., Dirden-Kramer, B., Mifflin, R. C., Reyes, V. E., Ryan, K. A., Elliott, S. N., Izumi, T., Boldogh, I., Mitra, S., Ernst, P. B., & Crowe, S. E. (2004). Helicobacter pylori and H2O2 increase AP endonuclease-1/redox factor-1 expression in human gastric epithelial cells. Gastroenterology, 127(3), 845-858. https://doi.org/10.1053/j.gastro.2004.06.017