1887

Abstract

To understand the biological processes within host cells induced by VacA, isogenic strains of (NCTC 11638 or 11638-Δ) were used to stimulate gastric cancer cells SGC7901, and differentially expressed genes in host cells were identified using cDNA microarray technology. More than 300 genes were found to alter their mRNA expression at different time points, among which 68 were related to the cytoskeleton, 87 were associated with cell cycle, cell death and proliferation, IL8 expression was also found to be up-regulated. Cells co-cultured with broth-culture supernatant (BCS) of NCTC 11638 showed more alteration in microtubule cytoskeleton morphology, as observed by laser scanning confocal microscopy, and a lower apoptosis rate, detected by flow cytometry, compared with those co-cultured with BCS of 11638-Δ. The supernatants of cells co-cultured with NCTC 11638 showed significantly higher IL8 expression than those co-cultured with 11638-Δ. It is concluded that VacA disrupts cytoskeletal architecture by influencing the expression of cytoskeleton-associated genes. VacA breaks the balance between cell proliferation and cell death by inducing the maladjustment of genes related to cell cycle. VacA is also able to induce the inflammatory response.

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2004-10-01
2024-04-18
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References

  1. Aoki M., Furusawa Y., Shibamoto Y., Kobayashi A., Tsujitani M. 2002; Effect of a hypoxic cell sensitizer doranidazole on the radiation-induced apoptosis of mouse L5178Y lymphoma cells. J Radiat Res (Tokyo) 43:161–166 [CrossRef]
    [Google Scholar]
  2. Bach S., Makristathis A., Rotter M., Hirschl A. M. 2002; Gene expression profiling in AGS cells stimulated with Helicobacter pylori isogenic strains ( cagA positive or cagA negative). Infect Immun 70:988–992 [CrossRef]
    [Google Scholar]
  3. Backert S., Schwarz T., Miehlke S. & 8 other authors; 2004; Functional analysis of the cag pathogenicity island in Helicobacter pylori isolates from patients with gastritis, peptic ulcer, and gastric cancer. Infect Immun 72:1043–1056 [CrossRef]
    [Google Scholar]
  4. Bagchi D., Bhattacharya G., Stohs S. J. 1996; Production of reactive oxygen species by gastric cells in association with Helicobacter pylori . Free Radic Res 24:439–450 [CrossRef]
    [Google Scholar]
  5. Bagchi D., McGinn T. R., Ye X., Bagchi M., Krohn R. L., Chatterjee A., Stohs S. J. 2002; Helicobacter pylori -induced oxidative stress and DNA damage in a primary culture of human gastric mucosal cells. Dig Dis Sci 47:1405–1412 [CrossRef]
    [Google Scholar]
  6. Bluhm W. F., Martin J. L., Mestril R., Dillmann W. H. 1998; Specific heat shock proteins protect microtubules during simulated ischemia in cardiac myocytes. Am J Physiol 275:H2243–H2249
    [Google Scholar]
  7. Chen G., Goeddel D. V. 2002; TNF-R1 signaling: a beautiful pathway. Science 296:1634–1635 [CrossRef]
    [Google Scholar]
  8. Chu Z. L., McKinsey T. A., Liu L., Gentry J. J., Malim M. H., Ballard D. W. 1997; Suppression of tumor necrosis factor-induced cell death by inhibitor of apoptosis c-IAP2 is under NF-κB control. Proc Natl Acad Sci U S A 94:10057–10062 [CrossRef]
    [Google Scholar]
  9. Crabtree J. E., Peichl P., Wyatt J. I., Stachl U., Lindley I. J. 1993; Gastric interleukin-8 and IgA IL-8 autoantibodies in Helicobacter pylori infection. Scand J Immunol 37:65–70 [CrossRef]
    [Google Scholar]
  10. Crabtree J. E., Wyatt J. I., Trejdosiewicz L. K., Peichl P., Nichols P. H., Ramsay N., Primrose J. N., Lindley I. J. 1994; Interleukin-8 expression in Helicobacter pylori infected, normal and neoplastic gastroduodenal mucosa. J Clin Pathol 47:61–66 [CrossRef]
    [Google Scholar]
  11. Crowe S. E., Alvarez L., Dytoc M., Hunt R. H., Muller M., Sherman P., Patel J., Jin Y., Ernst P. B. 1995; Expression of interleukin 8 and CD54 by human gastric epithelium after Helicobacter pylori infection in vitro. Gastroenterology 108:65–74 [CrossRef]
    [Google Scholar]
  12. Day A. S., Jones N. L., Policova Z., Jennings H. A., Yau E. K., Shannon P., Neumann A. W., Sherman P. M. 2001; Characterization of virulence factors of mouse-adapted Helicobacter pylori strain SS1 and effects on gastric hydrophobicity. Dig Dis Sci 46:1943–1951 [CrossRef]
    [Google Scholar]
  13. de Jonge R., Kusters J. G., Timmer M. S. & 8 other authors; 2001; The role of Helicobacter pylori virulence factors in interleukin production by monocytic cells. FEMS Microbiol Lett 196:235–238 [CrossRef]
    [Google Scholar]
  14. DeRisi J., Penland L., Brown P. O., Bittner M. L., Meltzer P. S., Ray M., Chen Y., Su Y. A., Trent J. M. 1996; Use of a cDNA microarray to analyse gene expression patterns in human cancer. Nat Genet 14:457–460 [CrossRef]
    [Google Scholar]
  15. Detweiler C. S., Cunanan D. B., Falkow S. 2001; Host microarray analysis reveals a role for the Salmonella response regulator phoP in human macrophage cell death. Proc Natl Acad Sci U S A 98:5850–5855 [CrossRef]
    [Google Scholar]
  16. Eaton K. A., Cover T. L., Tummuru M. K., Blaser M. J., Krakowka S. 1997; Role of vacuolating cytotoxin in gastritis due to Helicobacter pylori in gnotobiotic piglets. Infect Immun 65:3462–3464
    [Google Scholar]
  17. Eaton K. A., Kersulyte D., Mefford M., Danon S. J., Krakowka S., Berg D. E. 2001; Role of Helicobacter pylori cag region genes in colonization and gastritis in two animal models. Infect Immun 69:2902–2908 [CrossRef]
    [Google Scholar]
  18. Figura N., Guglielmetti P., Rossolini A., Barberi A., Cusi G., Musmanno R. A., Russi M., Quaranta S. 1989; Cytotoxin production by Campylobacter pylori strains isolated from patients with peptic ulcers and from patients with chronic gastritis only. J Clin Microbiol 27:225–226
    [Google Scholar]
  19. Fujiwara Y., Wyle F., Arakawa T., Domek M. J., Fukuda T., Kobayashi K., Tarnawski A. 1997; Helicobacter pylori culture supernatant inhibits binding and proliferative response of human gastric cells to epidermal growth factor: implications for H.pylori interference with ulcer healing?. Digestion 58:299–303 [CrossRef]
    [Google Scholar]
  20. Ghiara P., Marchetti M., Blaser M. J., Tummuru M. K., Cover T. L., Segal E. D., Tompkins L. S., Rappuoli R. 1995; Role of the Helicobacter pylori virulence factors vacuolating cytotoxin, CagA, and urease in a mouse model of disease. Infect Immun 63:4154–4160
    [Google Scholar]
  21. Go M. F., Cissell L., Graham D. Y. 1998; Failure to confirm association of vac A gene mosaicism with duodenal ulcer disease. Scand J Gastroenterol 33:132–136 [CrossRef]
    [Google Scholar]
  22. Goyal L. 2001; Cell death inhibition: keeping caspases in check. Cell 104:805–808 [CrossRef]
    [Google Scholar]
  23. Huang J., O'Toole P. W., Doig P., Trust T. J. 1995; Stimulation of interleukin-8 production in epithelial cell lines by Helicobacter pylori . Infect Immun 63:1732–1738
    [Google Scholar]
  24. Ito K., Hanazawa T., Tomita K., Barnes P. J., Adcock I. M. 2004; Oxidative stress reduces histone deacetylase 2 activity and enhances IL-8 gene expression: role of tyrosine nitration. Biochem Biophys Res Commun 315:240–245 [CrossRef]
    [Google Scholar]
  25. Keates S., Hitti Y. S., Upton M., Kelly C. P. 1997; Helicobacter pylori infection activates NF-κB in gastric epithelial cells. Gastroenterology 113:1099–1109 [CrossRef]
    [Google Scholar]
  26. Kim J. M., Kim J. S., Jung H. C., Song I. S., Kim C. Y. 2000; Virulence factors of Helicobacter pylori in Korean isolates do not influence proinflammatory cytokine gene expression and apoptosis in human gastric epithelial cells, nor do these factors influence the clinical outcome. J Gastroenterol 35:898–906 [CrossRef]
    [Google Scholar]
  27. Kodama K., Fujioka T., Ito A., Kubota T., Murakami K., Nasu M. 1996; Expression of vacuolating cytotoxin in clinical isolates of Helicobacter pylori . J Gastroenterol 31 (Suppl. 9):9–11
    [Google Scholar]
  28. Konturek S. J., Brzozowski T., Konturek P. C., Kwiecien S., Karczewska E., Drozdowicz D., Stachura J., Hahn E. G. 2000; Helicobacter pylori infection delays healing of ischaemia-reperfusion induced gastric ulcerations: new animal model for studying pathogenesis and therapy of H.pylori infection. Eur J Gastroenterol Hepatol 12:1299–1313 [CrossRef]
    [Google Scholar]
  29. Konturek J. W., Fischer H., Konturek P. C. & 9 other authors; 2001; Heat shock protein 70 (HSP70) in gastric adaptation to aspirin in Helicobacter pylori infection. J Physiol Pharmacol 52:153–164
    [Google Scholar]
  30. Kubota H., Hynes G., Carne A., Ashworth A., Willison K. 1994; Identification of six Tcp-1-related genes encoding divergent subunits of the TCP-1-containing chaperonin. Curr Biol 4:89–99 [CrossRef]
    [Google Scholar]
  31. Liao J., Lowthert L. A., Ghori N., Omary M. B. 1995; The 70-kDa heat shock proteins associate with glandular intermediate filaments in an ATP-dependent manner. J Biol Chem 270:915–922 [CrossRef]
    [Google Scholar]
  32. Macejak D. G., Luftig R. B. 1991; Stabilization of actin filaments at early times after adenovirus infection and in heat-shocked cells. Virus Res 19:31–45 [CrossRef]
    [Google Scholar]
  33. Maeda S., Yoshida H., Ogura K., Mitsuno Y., Hirata Y., Yamaji Y., Akanuma M., Shiratori Y., Omata M. 2000; H.pylori activates NF-κB through a signaling pathway involving IκB kinases, NF-κB-inducing kinase, TRAF2, and TRAF6 in gastric cancer cells. Gastroenterology 119:97–108 [CrossRef]
    [Google Scholar]
  34. Moss S. F., Calam J., Agarwal B., Wang S., Holt P. R. 1996; Induction of gastric epithelial apoptosis by Helicobacter pylori . Gut 38:498–501 [CrossRef]
    [Google Scholar]
  35. Naumann M., Wessler S., Bartsch C., Wieland B., Covacci A., Haas R., Meyer T. F. 1999; Activation of activator protein 1 and stress response kinases in epithelial cells colonized by Helicobacter pylori encoding the cag pathogenicity island. J Biol Chem 274:31655–31662 [CrossRef]
    [Google Scholar]
  36. Ogura K., Maeda S., Nakao M., Watanabe T., Tada M., Kyutoku T., Yoshida H., Shiratori Y., Omata M. 2000; Virulence factors of Helicobacter pylori responsible for gastric diseases in Mongolian gerbil. J Exp Med 192:1601–1610 [CrossRef]
    [Google Scholar]
  37. Pai R., Cover T. L., Tarnawski A. S. 1999; Helicobacter pylori vacuolating cytotoxin (VacA) disorganizes the cytoskeletal architecture of gastric epithelial cells. Biochem Biophys Res Commun 262:245–250 [CrossRef]
    [Google Scholar]
  38. Peek R. M. Jr, Miller G. G., Tham K. T., Perez-Perez G. I., Zhao X., Atherton J. C., Blaser M. J. 1995; Heightened inflammatory response and cytokine expression in vivo to cagA+ Helicobacter pylori strains. Lab Invest 73:760–770
    [Google Scholar]
  39. Peek R. M. Jr, Moss S. F., Tham K. T., Perez-Perez G. I., Wang S., Miller G. G., Atherton J. C., Holt P. R., Blaser M. J. 1997; Helicobacter pylori cagA+ strains and dissociation of gastric epithelial cell proliferation from apoptosis. J Natl Cancer Inst 89:863–868 [CrossRef]
    [Google Scholar]
  40. Rudi J., Kuck D., Strand S., von Herbay A., Mariani S. M., Krammer P. H., Galle P. R., Stremmel W. 1998; Involvement of the CD95 (APO-1/Fas) receptor and ligand system in Helicobacter pylori -induced gastric epithelial apoptosis. J Clin Invest 102:1506–1514 [CrossRef]
    [Google Scholar]
  41. Schreck R., Albermann K., Baeuerle P. A. 1992; Nuclear factor kappa B: an oxidative stress-responsive transcription factor of eukaryotic cells (a review). Free Radic Res Commun 17:221–237 [CrossRef]
    [Google Scholar]
  42. Shimada T., Watanabe N., Hiraishi H., Terano A. 1999; Redox regulation of interleukin-8 expression in MKN28 cells. Dig Dis Sci 44:266–273 [CrossRef]
    [Google Scholar]
  43. Straubinger R. K., Greiter A., McDonough S. P. & 7 other authors; 2003; Quantitative evaluation of inflammatory and immune responses in the early stages of chronic Helicobacter pylori infection. Infect Immun 71:2693–2703 [CrossRef]
    [Google Scholar]
  44. Telford J. L., Ghiara P., Dell'Orco M. & 11 other authors; 1994; Gene structure of the Helicobacter pylori cytotoxin and evidence of its key role in gastric disease. J Exp Med 179:1653–1658 [CrossRef]
    [Google Scholar]
  45. Trush M. A., Kensler T. W. 1991; An overview of the relationship between oxidative stress and chemical carcinogenesis. Free Radic Biol Med 10:201–209 [CrossRef]
    [Google Scholar]
  46. van Doorn N. E., Namavar F., Sparrius M., Stoof J., van Rees E. P., van Doorn L. J., Vandenbroucke-Grauls C. M. 1999; Helicobacter pylori -associated gastritis in mice is host and strain specific. Infect Immun 67:3040–3046
    [Google Scholar]
  47. van Geel M., van Deutekom J. C., van Staalduinen A. & 7 other authors; 2000; Identification of a novel β-tubulin subfamily with one member (TUBB4Q) located near the telomere of chromosome region 4q35. Cytogenet Cell Genet 88:316–321 [CrossRef]
    [Google Scholar]
  48. von Herbay A., Rudi J. 2000; Role of apoptosis in gastric epithelial turnover. Microsc Res Tech 48:303–311 [CrossRef]
    [Google Scholar]
  49. Wagner S., Beil W., Westermann J., Logan R. P., Bock C. T., Trautwein C., Bleck J. S., Manns M. P. 1997; Regulation of gastric epithelial cell growth by Helicobacter pylori : offdence for a major role of apoptosis. Gastroenterology 113:1836–1847 [CrossRef]
    [Google Scholar]
  50. Winrow V. R., Winyard P. G., Morris C. J., Blake D. R. 1993; Free radicals in inflammation: second messengers and mediators of tissue destruction. Br Med Bull 49:506–522
    [Google Scholar]
  51. Xia H. H., Talley N. J. 2001; Apoptosis in gastric epithelium induced by Helicobacter pylori infection: implications in gastric carcinogenesis. Am J Gastroenterol 96:16–26
    [Google Scholar]
  52. Yuan J. P., Li T., Shi X. D., Hu B. Y., Yang G. Z., Tong S. Q., Guo X. K. 2003a; Deletion of Helicobacter pylori vacuolating cytotoxin gene by introduction of directed mutagenesis. World J Gastroenterol 9:2251–2257
    [Google Scholar]
  53. Yuan J. P., Zhao W., Wang H. T., Wu K. Y., Li T., Guo X. K., Tong S. Q. 2003b; Coxsackievirus B3-induced apoptosis and caspase-3. Cell Res 13:203–209 [CrossRef]
    [Google Scholar]
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