Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury

Duane T. Smoot, Tollie B. Elliott, Hein W. Verspaget, Dana Jones, Cornell R. Allen, Kurt G. Vernon, Theodore Bremner, La Creis R Kidd, Kyung S. Kim, John D. Groupman, Hassan Ashktorab

Research output: Contribution to journalArticle

Abstract

Risk factors for gastric cancer are receiving renewed attention in light of the recent positive association of Helicobacter pylori infection with gastric cancer. The effect of H.pylori on the balance between oxidants and anti-oxidants in the stomach is not well known. In this study, we investigated if exposure of gastric cells to H.pylori increases oxidant-associated gastric epithelial cell injury. A human gastric epithelial cell line (AGS) was grown on 96-well clusters, then exposed overnight to either live H.pylori (four cagA+ and four cagA-) or broth culture supernatant from an isogenic H.pylori cagA+ strain with and without vacA activity. Incubation of AGS cells with cagA+ and cagA- H.pylori strains before exposure to reactive oxygen species (ROS) reduced cell viability on average to 73.7% and 39.5% of controls, respectively. The percent viability of cells exposed to ROS after incubation with control broth, vacA- broth and vacA+ broth was 97.7%, 70.5% and 63.5%, respectively. Experiments were then performed to evaluate the effects of H.pylori exposure on the activities of ROS-scavenging enzymes [catalase, glutathione peroxidase and superoxide dismutase (SOD)] and formation of 8-hydroxy-2-deoxyguanosine (8-OH-dG) adducts in AGS cells. Overnight exposure to cagA- strains reduced catalase activity by 42%; in contrast, exposure to cagA+ H.pylori strains increased catalase activity by 51%. Glutathione peroxidase activity increased with exposure to both cagA- and cagA+ strains by 95% and 240%, respectively. Total SOD activity increased 156% after exposure to cagA+ strains and was marginally increased (52%) with exposure to cagA- strains. CuZn-SOD protein levels, assayed by enzyme-linked immunosorbent assay, were not significantly altered by exposure to H.pylori strains; however, Mn-SOD concentrations were significantly increased (P <0.02) after exposure to both cagA- and cagA+ H.pylori strains. Exposure of AGS cells to cagA+ and cagA- H.pylori was associated with, on average, 44.5 and 99.0 8-OH-dG/106 dG, respectively. The increase in catalase, glutathione peroxidase and SOD activity is associated with fewer 8-OH-dG DNA adducts and reduced susceptibility of AGS cells to lethal injury from ROS after exposure to cagA+ H.pylori strains when compared with exposure to cagA- H.pylori strains. Alteration in the activity of ROS-scavenging enzymes by the presence of H.pylori may in part be responsible for the increased risk of gastric cancer in persons infected with H.pylori.

Original languageEnglish (US)
Pages (from-to)2091-2095
Number of pages5
JournalCarcinogenesis
Volume21
Issue number11
StatePublished - 2000
Externally publishedYes

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Pylorus
Helicobacter pylori
Stomach
Epithelial Cells
Oxygen
Wounds and Injuries
Superoxide Dismutase
Reactive Oxygen Species
Catalase
Glutathione Peroxidase
Oxidants
Stomach Neoplasms
Cell Survival
DNA Adducts
Helicobacter Infections
Enzymes

ASJC Scopus subject areas

  • Cancer Research

Cite this

Smoot, D. T., Elliott, T. B., Verspaget, H. W., Jones, D., Allen, C. R., Vernon, K. G., ... Ashktorab, H. (2000). Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury. Carcinogenesis, 21(11), 2091-2095.

Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury. / Smoot, Duane T.; Elliott, Tollie B.; Verspaget, Hein W.; Jones, Dana; Allen, Cornell R.; Vernon, Kurt G.; Bremner, Theodore; Kidd, La Creis R; Kim, Kyung S.; Groupman, John D.; Ashktorab, Hassan.

In: Carcinogenesis, Vol. 21, No. 11, 2000, p. 2091-2095.

Research output: Contribution to journalArticle

Smoot, DT, Elliott, TB, Verspaget, HW, Jones, D, Allen, CR, Vernon, KG, Bremner, T, Kidd, LCR, Kim, KS, Groupman, JD & Ashktorab, H 2000, 'Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury', Carcinogenesis, vol. 21, no. 11, pp. 2091-2095.
Smoot DT, Elliott TB, Verspaget HW, Jones D, Allen CR, Vernon KG et al. Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury. Carcinogenesis. 2000;21(11):2091-2095.
Smoot, Duane T. ; Elliott, Tollie B. ; Verspaget, Hein W. ; Jones, Dana ; Allen, Cornell R. ; Vernon, Kurt G. ; Bremner, Theodore ; Kidd, La Creis R ; Kim, Kyung S. ; Groupman, John D. ; Ashktorab, Hassan. / Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury. In: Carcinogenesis. 2000 ; Vol. 21, No. 11. pp. 2091-2095.
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abstract = "Risk factors for gastric cancer are receiving renewed attention in light of the recent positive association of Helicobacter pylori infection with gastric cancer. The effect of H.pylori on the balance between oxidants and anti-oxidants in the stomach is not well known. In this study, we investigated if exposure of gastric cells to H.pylori increases oxidant-associated gastric epithelial cell injury. A human gastric epithelial cell line (AGS) was grown on 96-well clusters, then exposed overnight to either live H.pylori (four cagA+ and four cagA-) or broth culture supernatant from an isogenic H.pylori cagA+ strain with and without vacA activity. Incubation of AGS cells with cagA+ and cagA- H.pylori strains before exposure to reactive oxygen species (ROS) reduced cell viability on average to 73.7{\%} and 39.5{\%} of controls, respectively. The percent viability of cells exposed to ROS after incubation with control broth, vacA- broth and vacA+ broth was 97.7{\%}, 70.5{\%} and 63.5{\%}, respectively. Experiments were then performed to evaluate the effects of H.pylori exposure on the activities of ROS-scavenging enzymes [catalase, glutathione peroxidase and superoxide dismutase (SOD)] and formation of 8-hydroxy-2-deoxyguanosine (8-OH-dG) adducts in AGS cells. Overnight exposure to cagA- strains reduced catalase activity by 42{\%}; in contrast, exposure to cagA+ H.pylori strains increased catalase activity by 51{\%}. Glutathione peroxidase activity increased with exposure to both cagA- and cagA+ strains by 95{\%} and 240{\%}, respectively. Total SOD activity increased 156{\%} after exposure to cagA+ strains and was marginally increased (52{\%}) with exposure to cagA- strains. CuZn-SOD protein levels, assayed by enzyme-linked immunosorbent assay, were not significantly altered by exposure to H.pylori strains; however, Mn-SOD concentrations were significantly increased (P <0.02) after exposure to both cagA- and cagA+ H.pylori strains. Exposure of AGS cells to cagA+ and cagA- H.pylori was associated with, on average, 44.5 and 99.0 8-OH-dG/106 dG, respectively. The increase in catalase, glutathione peroxidase and SOD activity is associated with fewer 8-OH-dG DNA adducts and reduced susceptibility of AGS cells to lethal injury from ROS after exposure to cagA+ H.pylori strains when compared with exposure to cagA- H.pylori strains. Alteration in the activity of ROS-scavenging enzymes by the presence of H.pylori may in part be responsible for the increased risk of gastric cancer in persons infected with H.pylori.",
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AU - Smoot, Duane T.

AU - Elliott, Tollie B.

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AU - Jones, Dana

AU - Allen, Cornell R.

AU - Vernon, Kurt G.

AU - Bremner, Theodore

AU - Kidd, La Creis R

AU - Kim, Kyung S.

AU - Groupman, John D.

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N2 - Risk factors for gastric cancer are receiving renewed attention in light of the recent positive association of Helicobacter pylori infection with gastric cancer. The effect of H.pylori on the balance between oxidants and anti-oxidants in the stomach is not well known. In this study, we investigated if exposure of gastric cells to H.pylori increases oxidant-associated gastric epithelial cell injury. A human gastric epithelial cell line (AGS) was grown on 96-well clusters, then exposed overnight to either live H.pylori (four cagA+ and four cagA-) or broth culture supernatant from an isogenic H.pylori cagA+ strain with and without vacA activity. Incubation of AGS cells with cagA+ and cagA- H.pylori strains before exposure to reactive oxygen species (ROS) reduced cell viability on average to 73.7% and 39.5% of controls, respectively. The percent viability of cells exposed to ROS after incubation with control broth, vacA- broth and vacA+ broth was 97.7%, 70.5% and 63.5%, respectively. Experiments were then performed to evaluate the effects of H.pylori exposure on the activities of ROS-scavenging enzymes [catalase, glutathione peroxidase and superoxide dismutase (SOD)] and formation of 8-hydroxy-2-deoxyguanosine (8-OH-dG) adducts in AGS cells. Overnight exposure to cagA- strains reduced catalase activity by 42%; in contrast, exposure to cagA+ H.pylori strains increased catalase activity by 51%. Glutathione peroxidase activity increased with exposure to both cagA- and cagA+ strains by 95% and 240%, respectively. Total SOD activity increased 156% after exposure to cagA+ strains and was marginally increased (52%) with exposure to cagA- strains. CuZn-SOD protein levels, assayed by enzyme-linked immunosorbent assay, were not significantly altered by exposure to H.pylori strains; however, Mn-SOD concentrations were significantly increased (P <0.02) after exposure to both cagA- and cagA+ H.pylori strains. Exposure of AGS cells to cagA+ and cagA- H.pylori was associated with, on average, 44.5 and 99.0 8-OH-dG/106 dG, respectively. The increase in catalase, glutathione peroxidase and SOD activity is associated with fewer 8-OH-dG DNA adducts and reduced susceptibility of AGS cells to lethal injury from ROS after exposure to cagA+ H.pylori strains when compared with exposure to cagA- H.pylori strains. Alteration in the activity of ROS-scavenging enzymes by the presence of H.pylori may in part be responsible for the increased risk of gastric cancer in persons infected with H.pylori.

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