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Pathology & Oncology Research

, Volume 20, Issue 4, pp 839–846 | Cite as

Oxidative DNA Damage as a Potential Early Biomarker of Helicobacter pylori Associated Carcinogenesis

  • Yasir Raza
  • Adnan Khan
  • Amber Farooqui
  • Muhammad MubarakEmail author
  • Alex Facista
  • Syed Shakeel Akhtar
  • Saeed Khan
  • Javed Iqbal Kazi
  • Carol Bernstein
  • Shahana Urooj Kazmi
Research

Abstract

Helicobacter pylori infection is an established risk factor for gastritis, gastric ulcer, peptic ulcer and gastric cancer. CagA +ve H. pylori has been associated with oxidative DNA damage of gastric mucosa but their combined role in the development of gastric cancer is still unknown. Here we compare the combined expression of cagA and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in normal, gastritis and gastric cancer tissues. Two hundred gastric biopsies from patients with dyspeptic symptoms, 70 gastric cancer tissue samples and 30 gastric biopsies from non-dyspeptic individuals (controls) were included in this study and 8-OHdG was detected by immunohistochemistry (IHC). Histological features and the presence of H. pylori infection were demonstrated by Hematoxylin and Eosin (HE), Giemsa and alcian blue-periodic acid-Schiff ± diastase (AB-PAS ± D) staining. DNA was extracted from tissues and polymerase chain reaction (PCR) performed to determine the presence of ureaseA and cagA genes of H. pylori. The results showed the presence of H. pylori in 106 (53 %) gastric biopsies out of 200 dyspeptic patients, including 70 (66 %) cases of cagA + ve H. pylori. The presence of cagA gene and high expression of 8-OHdG was highly correlated with severe gastric inflammation and gastric cancer particularly, in cases with infiltration of chronic inflammatory cells (36.8 % cagA + ve, 18 %), neutrophilic activity (47.2 %, 25.5 %), intestinal metaplasia (77.7 %, 35.7 %) and intestinal type gastric cancer (95 %, 95.4 %) (p ≤ 0.01). In conclusion, H. Pylori cagA gene expression and the detection of 8-OHdG adducts in gastric epithelium can serve as potential early biomarkers of H. Pylori-associated gastric carcinogenesis.

Keywords

Helicobacter pylori Gastritis 8-hydroxy-2′-deoxyguanosine Gastric cancer Immunohistochemistry 

Notes

Acknowledgments

We are also highly thankful to Mr. Nazim, Department of Histopathology, Zaiuddun University and Hospital, for his assistance in collecting samples. We are also highly grateful to Mrs. Nikki Kelvin for the technical revision of manuscript and Dr Saleem Iqbal for helping in statistical analysis.

Financial disclosures

Funds to conduct study were provided by HEC, Pakistan. Dr. Bernstein provided consumables and bench space at University of Arizona, USA. No other financial declarations by any of the authors.

Conflict of interest

None by any of the authors.

References

  1. 1.
    Auten RL, Davis JM (2009) Oxygen toxicity and reactive oxygen species: the devil is in the details. Pediatr Res 66:121–127PubMedCrossRefGoogle Scholar
  2. 2.
    Klaunig JE, Kamendulis LM, Hocevar BA (2010) Oxidative stress and oxidative damage in carcinogenesis. Toxicol Pathol 38:96–109PubMedCrossRefGoogle Scholar
  3. 3.
    Inokuma T, Haraguchi M, Fujita F, Tajima Y, Kanematsu T (2009) Oxidative stress and tumor progression in colorectal cancer. Hepatogastroenterology 56:343–347PubMedGoogle Scholar
  4. 4.
    Smoot DT, Elliott TB, Verspaget HW, Jones D, Allen CR, Vernon KG et al (2000) Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury. Carcinogenesis 21:2091–2095PubMedCrossRefGoogle Scholar
  5. 5.
    Farinati F, Cardin R, Russo VM, Busatto G, Franco M, Rugge M (2003) Helicobacter pylori CagA status, mucosal oxidative damage and gastritis phenotype: a potential pathway to cancer? Helicobacter 8:227–234PubMedCrossRefGoogle Scholar
  6. 6.
    Varon C, Mosnier JF, Lehours P, Matysiak-Budnik T, Mégraud F (2009) Gastric carcinogenesis and Helicobacter pylori infection. Methods Mol Biol 511:237–265PubMedCrossRefGoogle Scholar
  7. 7.
    Valavanidis A, Vlachogianni T, Fiotakis C (2009) 8-hydroxy-2′-deoxyguanosine (8-OHdG): a critical biomarker of oxidative stress and carcinogenesis. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 27:120–139PubMedCrossRefGoogle Scholar
  8. 8.
    Kuchino Y, Mori F, Kasai H, Inoue H, Iwai S, Miura K et al (1987) Misreading of DNA templates containing 8-hydroxydeoxyguanosine at the modified base and at adjacent residues. Nature 327:77–79PubMedCrossRefGoogle Scholar
  9. 9.
    Lee SF, Pervaiz S (2011) Assessment of oxidative stress-induced DNA damage by immunoflourescent analysis of 8-OxodG. Methods Cell Biol 103:99–113PubMedCrossRefGoogle Scholar
  10. 10.
    Møller H, Heseltine E, Vainio H (1995) Working group report on schistosomes, liver flukes and Helicobacter pylori. Int J Cancer 60:587–589PubMedCrossRefGoogle Scholar
  11. 11.
    Marshall BJ, Warren JR (1984) Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1:1311–1315PubMedCrossRefGoogle Scholar
  12. 12.
    Yamaoka Y (2008) Increasing evidence of the role of Helicobacter pylori SabA in the pathogenesis of gastroduodenal disease. J Infect Dev Ctries 2:174–181PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    Asahi M, Azuma T, Ito S, Ito Y, Suto H, Nagai Y et al (2000) Helicobacter pylori CagA protein can be tyrosine phosphorylated in gastric epithelial cells. J Exp Med 191:593–602PubMedCrossRefPubMedCentralGoogle Scholar
  14. 14.
    Odenbreit S, Püls J, Sedlmaier B, Gerland E, Fischer W, Haas R (2000) Translocation of Helicobacter pylori CagA into gastric epithelial cells by type IV secretion. Science 287:1497–1500PubMedCrossRefGoogle Scholar
  15. 15.
    Danese S, Cremonini F, Armuzzi A, Candelli M, Papa A, Ojetti V et al (2001) Helicobacter pylori CagA-positive strains affect oxygen free radicals generation by gastric mucosa. Scand J Gastroenterol 36:247–250PubMedCrossRefGoogle Scholar
  16. 16.
    Suzuki H, Suzuki M, Imaeda H, Hibi T (2009) Helicobacter pylori and microcirculation. Microcirculation 16:547–558PubMedCrossRefGoogle Scholar
  17. 17.
    González CA, Figueiredo C, Lic CB, Ferreira RM, Pardo ML, Ruiz Liso JM et al (2011) Helicobacter pylori cagA and vacA genotypes as predictors of progression of gastric preneoplastic lesions: a long-term follow-up in a high-risk area in Spain. Am J Gastroenterol 106:867–874PubMedCrossRefGoogle Scholar
  18. 18.
    Blaser MJ, Perez-Perez GI, Kleanthous H, Cover TL, Peek RM, Chyou PH et al (1995) Infection with Helicobacter pylori strains possessing cagA is associated with an increased risk of developing adenocarcinoma of the stomach. Cancer Res 55:2111–2115PubMedGoogle Scholar
  19. 19.
    Singh K, Ghoshal UC (2006) Causal role of Helicobacter pylori infection in gastric cancer: an Asian enigma. World J Gastroenterol 12:1346–1351PubMedPubMedCentralGoogle Scholar
  20. 20.
    Qureshi TZ, Bilal R, Saleem K, Zafar S (2008) General prevalence of Helicobacter pylori infection in dyspeptic population of Islamabad, Pakistan. Nucleus 45:157–162Google Scholar
  21. 21.
    Taj Y, Essa F, Kazmi SU, Abdullah E (2003) Sensitivity and specificity of various diagnostic tests in the detection of Helicobacter pylori. J Coll Physicians Surg Pak 13:90–93PubMedGoogle Scholar
  22. 22.
    Khan SS, Zulfiqar A, Danish KF, Sauwal M, Bashir S, Zaman S (2008) Prevalence of H. Pylori infection in patients with gastroduodenal disease in Pakistan. Rawal Med J 33:1–6Google Scholar
  23. 23.
    Yakoob J, Jafri W, Jafri N, Islam M, Abid S, Hamid S et al (2005) Prevalence of non-Helicobacter pylori duodenal ulcer in Karachi, Pakistan. World J Gastroenterol 11:3562–3565PubMedGoogle Scholar
  24. 24.
    Peek RM Jr, Blaser MJ (2002) Helicobacter pylori and gastrointestinal tract adenocarcinomas. Nat Rev Cancer 2:28–37PubMedCrossRefGoogle Scholar
  25. 25.
    Dixon MF, Genta RM, Yardley JH, Correa P (1996) Classification and grading of gastritis. The updated Sydney System. International Workshop on the Histopathology of Gastritis, Houston 1994. Am J Surg Pathol 20:1161–1181PubMedCrossRefGoogle Scholar
  26. 26.
    Khan S, Jaffer NN, Khan MN, Rai MA, Shafiq M, Ali A et al (2007) Human papillomavirus subtype 16 is common in Pakistani women with cervical carcinoma. Int J Infect Dis 11:313–317PubMedCrossRefGoogle Scholar
  27. 27.
    Ashton-Key M, Diss TC, Isaacson PG (1996) Detection of Helicobacter pylori in gastric biopsy and resection specimens. J Clin Pathol 49:107–111PubMedCrossRefPubMedCentralGoogle Scholar
  28. 28.
    Qabandi AA, Mustafa AS, Siddique I, Khajah AK, Madda JP, Junaid TA (2005) Distribution of vacA and cagA genotypes of Helicobacter pylori in Kuwait. Acta Trop 93:283–288PubMedCrossRefGoogle Scholar
  29. 29.
    Papa A, Danese S, Sgambato A, Ardito R, Zannoni G, Rinelli A et al (2002) Role of Helicobacter pylori CagA +infection in determining oxidative DNA damage in gastric mucosa. Scand J Gastroenterol 37:409–413PubMedCrossRefGoogle Scholar
  30. 30.
    Rasheed F, Khan A, Kazmi SU (2009) Bacteriological analysis, antimicrobial susceptibility and detection of 16S rRNA gene of Helicobacter pylori by PCR in drinking water samples of earthquake affected areas and other parts of Pakistan. Malay J Microbiol 5:123–127Google Scholar
  31. 31.
    Gao L, Michel A, Weck MN, Arndt V, Pawlita M, Brenner H (2009) Helicobacter pylori infection and gastric cancer risk: evaluation of 15H. pylori proteins determined by novel multiplex serology. Cancer Res 69:6164–6170PubMedCrossRefGoogle Scholar
  32. 32.
    Kido M, Watanabe N, Aoki N, Iwamoto S, Nishiura H, Maruoka R et al (2011) Dual roles of CagA protein in Helicobacter pylori-induced chronic gastritis in mice. Biochem Biophys Res Commun 412:266–272PubMedCrossRefGoogle Scholar
  33. 33.
    Hatakeyama M (2009) Helicobacter pylori and gastric carcinogenesis. J Gastroenterol 44:239–248PubMedCrossRefGoogle Scholar
  34. 34.
    Konturek PC, Konturek SJ, Brzozowski T (2009) Helicobacter pylori infection in gastric cancerogenesis. J Physiol Pharmacol 60:3–21PubMedGoogle Scholar
  35. 35.
    Davies GR, Simmonds NJ, Stevens TR, Sheaff MT, Banatvala N, Laurenson IF et al (1994) Helicobacter pylori stimulates antral mucosal reactive oxygen metabolite production in vivo. Gut 35:179–185PubMedCrossRefPubMedCentralGoogle Scholar
  36. 36.
    Farinati F, Cardin R, Cassaro M, Bortolami M, Nitti D, Tieppo C et al (2008) Helicobacter pylori, inflammation, oxidative damage and gastric cancer: a morphological, biological and molecular pathway. Eur J Cancer Prev 17:195–200PubMedCrossRefGoogle Scholar
  37. 37.
    Katsurahara M, Kobayashi Y, Iwasa M, Ma N, Inoue H, Fujita N et al (2009) Reactive nitrogen species mediate DNA damage in Helicobacter pylori-infected gastric mucosa. Helicobacter 14:552–558PubMedCrossRefGoogle Scholar
  38. 38.
    Everett SM, White KL, Drake IM, Schorah CJ, Calvert RJ, Skinner C et al (2002) The effect of Helicobacter pylori infection on levels of DNA damage in gastric epithelial cells. Helicobacter 7:271–280PubMedCrossRefGoogle Scholar
  39. 39.
    Wroblewski LE, Peek RM Jr, Wilson KT (2010) Helicobacter pylori and gastric cancer: factors that modulate disease risk. Clin Microbiol Rev 23:713–739PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2014

Authors and Affiliations

  • Yasir Raza
    • 1
    • 2
  • Adnan Khan
    • 1
  • Amber Farooqui
    • 1
    • 6
  • Muhammad Mubarak
    • 3
    Email author
  • Alex Facista
    • 2
  • Syed Shakeel Akhtar
    • 4
  • Saeed Khan
    • 1
  • Javed Iqbal Kazi
    • 3
    • 5
  • Carol Bernstein
    • 2
  • Shahana Urooj Kazmi
    • 1
  1. 1.Immunology and Infectious Diseases Research Laboratory, Department of MicrobiologyUniversity of KarachiKarachiPakistan
  2. 2.Department of Cellular and Molecular Medicine, College of MedicineUniversity of ArizonaPhoenixUSA
  3. 3.Department of HistopathologySindh Institute of Urology and TransplantationKarachiPakistan
  4. 4.Department of Surgery and MedicineCivil HospitalKarachiPakistan
  5. 5.Department of HistopathologyZiauddin University and HospitalKarachiPakistan
  6. 6.Division of Immunology, International Institute of Infection and ImmunityShantou University Medical CollegeShantouChina

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