Digestive Diseases and Sciences

, Volume 58, Issue 1, pp 140–149 | Cite as

H. pylori Infection Is Associated with DNA Damage of Lgr5-Positive Epithelial Stem Cells in the Stomach of Patients with Gastric Cancer

  • Takeshi Uehara
  • Deqin Ma
  • Yuan Yao
  • John P. Lynch
  • Knashawn Morales
  • Amy Ziober
  • Michael Feldman
  • Hiroyoshi Ota
  • Antonia R. Sepulveda
Original Article



H. pylori (Hp) infection is a major risk factor in gastric carcinogenesis leading to epithelial mutagenesis, and may affect gastric epithelial stem cells.


To characterize the expression of Lgr5, a marker of epithelial stem cells in human gastric mucosa, to determine whether Hp infection affects Lgr5-positive epithelial cells (LPECs) and whether LPECs are susceptible to DNA damage associated with Hp infection.


Lgr5 expression was characterized in non-neoplastic gastric mucosa from 52 patients (34 with and 18 without gastric cancer (GC); 21 Hp-positive (Hp+) and 31 Hp-negative (Hp−)) by immunohistochemical and immunofluorescence staining. To determine the extent of DNA damage in LPECs, nuclear 8-hydroxydeoxyguanosine (8OHdG), a marker of DNA damage associated with oxidative stress, was measured by quantitative spectral image analysis.


LPECs were primarily present in gastric antrum. Higher numbers of LPECs were seen in Hp+ than in Hp− non-neoplastic mucosa of GC patients, P = .006, but not in patients without GC. 8OHdG levels in LPECs were significantly higher than in Lgr5-negative epithelial cells in Hp+ GC patients (P = .012) but not in Hp− cases (P = .414), whereas no difference was seen between Hp+ and Hp− mucosa of patients without GC.


The Lgr5-positive epithelial stem cell pool is expanded in Hp-associated gastritis in the antrum of patients with GC. In GC patients with active Hp infection, LPECs may be more susceptible to DNA damage than Lgr5-negative epithelial cells, suggesting that Hp infection may contribute to GC risk by affecting epithelial stem cells in the human stomach.


Helicobacter pylori Gastric DNA damage 8OHdG Lgr5 Stem cells 


Conflict of interest


Supplementary material

10620_2012_2360_MOESM1_ESM.docx (29 kb)
Supplementary material 1 (DOCX 29 kb)
10620_2012_2360_MOESM2_ESM.tif (9.8 mb)
Supplementary material 2 (TIFF 10023 kb)
10620_2012_2360_MOESM3_ESM.tif (24.5 mb)
Supplementary material 3 (TIFF 25119 kb)
10620_2012_2360_MOESM4_ESM.tif (24.8 mb)
Supplementary material 4 (TIFF 25390 kb)
10620_2012_2360_MOESM5_ESM.tif (6.7 mb)
Supplementary material 5 (TIFF 6833 kb)
10620_2012_2360_MOESM6_ESM.tif (8 mb)
Supplementary material 6 (TIFF 8158 kb)
10620_2012_2360_MOESM7_ESM.tif (7 mb)
Supplementary material 7 (TIFF 7180 kb)
10620_2012_2360_MOESM8_ESM.tif (31.6 mb)
Supplementary material 8 (TIFF 32346 kb)
10620_2012_2360_MOESM9_ESM.docx (35 kb)
Supplementary material 9 (DOCX 35 kb)


  1. 1.
    Warren JR, Marshall B. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1983;1:1273–1275.Google Scholar
  2. 2.
    Marshall BJ. Helicobacter pylori: the etiologic agent for peptic ulcer. JAMA. 1995;274:1064–1066.PubMedCrossRefGoogle Scholar
  3. 3.
    Kimura K. Gastritis and gastric cancer. Asia. Gastroenterol Clin North Am. 2000;29:609–621.PubMedCrossRefGoogle Scholar
  4. 4.
    Dixon MF, Genta RM, Yardley JH, Correa P. Classification and grading of gastritis. The updated Sydney system. International workshop on the histopathology of gastritis, Houston 1994. Am J Surg Pathol. 1996;20:1161–1181.PubMedCrossRefGoogle Scholar
  5. 5.
    Herrera V, Parsonnet J. Helicobacter pylori and gastric adenocarcinoma. Clin Microbiol Infect. 2009;15:971–976.PubMedCrossRefGoogle Scholar
  6. 6.
    Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010;127:2893–2917.PubMedCrossRefGoogle Scholar
  7. 7.
    Kim JJ, Tao H, Carloni E, Leung WK, Graham DY, Sepulveda AR. Helicobacter pylori impairs DNA mismatch repair in gastric epithelial cells. Gastroenterology. 2002;123:542–553.PubMedCrossRefGoogle Scholar
  8. 8.
    Yao Y, Tao H, Park DI, Sepulveda JL, Sepulveda AR. Demonstration and characterization of mutations induced by Helicobacter pylori organisms in gastric epithelial cells. Helicobacter. 2006;11:272–286.PubMedCrossRefGoogle Scholar
  9. 9.
    Sepulveda AR, Yao Y, Yan W, et al. CpG methylation and reduced expression of O6-methylguanine DNA methyltransferase is associated with Helicobacter pylori infection. Gastroenterology. 2010;138:1836–1844.PubMedCrossRefGoogle Scholar
  10. 10.
    Qiao XT, Gumucio DL. Current molecular markers for gastric progenitor cells and gastric cancer stem cells. J Gastroenterol. 2011;46:855–865.PubMedCrossRefGoogle Scholar
  11. 11.
    Barker N, Huch M, Kujala P, et al. Lgr5(+ve) stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. Cell Stem Cell. 2010;6:25–36.PubMedCrossRefGoogle Scholar
  12. 12.
    Simon E, Petke D, Boger C, et al. The spatial distribution of LGR5+ cells correlates with gastric cancer progression. PLoS ONE. 2012;7:e35486.PubMedCrossRefGoogle Scholar
  13. 13.
    Barker N, van Es JH, Kuipers J, et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature. 2007;449:1003–1007.PubMedCrossRefGoogle Scholar
  14. 14.
    Becker L, Huang Q, Mashimo H. Immunostaining of Lgr5, an intestinal stem cell marker, in normal and premalignant human gastrointestinal tissue. Sci World J. 2008;8:1168–1176.CrossRefGoogle Scholar
  15. 15.
    Farinati F, Cardin R, Degan P, et al. Oxidative DNA damage accumulation in gastric carcinogenesis. Gut. 1998;42:351–356.PubMedCrossRefGoogle Scholar
  16. 16.
    Hahm KB, Lee KJ, Choi SY, et al. Possibility of chemoprevention by the eradication of Helicobacter pylori: oxidative DNA damage and apoptosis in H. pylori infection. Am J Gastroenterol. 1997;92:1853–1857.PubMedGoogle Scholar
  17. 17.
    Kuchino Y, Mori F, Kasai H, et al. Misreading of DNA templates containing 8-hydroxydeoxyguanosine at the modified base and at adjacent residues. Nature. 1987;327:77–79.PubMedCrossRefGoogle Scholar
  18. 18.
    Barker N, Clevers H. Leucine-rich repeat-containing G-protein-coupled receptors as markers of adult stem cells. Gastroenterology. 2010;138:1681–1696.PubMedCrossRefGoogle Scholar
  19. 19.
    Yao Y, Tao H, Kim JJ, et al. Alterations of DNA mismatch repair proteins and microsatellite instability levels in gastric cancer cell lines. Lab Invest. 2004;84:915–922.PubMedCrossRefGoogle Scholar
  20. 20.
    Yoshizawa K, Jelezcova E, Brown AR, et al. Gastrointestinal hyperplasia with altered expression of DNA polymerase beta. PLoS ONE. 2009;4:e6493.PubMedCrossRefGoogle Scholar
  21. 21.
    Houghton J, Stoicov C, Nomura S, et al. Gastric cancer originating from bone marrow-derived cells. Science. 2004;306:1568–1571.PubMedCrossRefGoogle Scholar
  22. 22.
    Correa P, Houghton J. Carcinogenesis of Helicobacter pylori. Gastroenterology. 2007;133:659–672.PubMedCrossRefGoogle Scholar
  23. 23.
    Mills JC, Shivdasani RA. Gastric epithelial stem cells. Gastroenterology. 2011;140:412–424.PubMedCrossRefGoogle Scholar
  24. 24.
    Correa P. Human gastric carcinogenesis: a multistep and multifactorial process—First American Cancer Society Award Lecture on Cancer Epidemiology and Prevention. Cancer Res. 1992;52:6735–6740.PubMedGoogle Scholar
  25. 25.
    Gologan A, Graham DY, Sepulveda AR. Molecular markers in Helicobacter pylori-associated gastric carcinogenesis. Clin Lab Med. 2005;25:197–222.PubMedCrossRefGoogle Scholar
  26. 26.
    Farinati F, Cardin R, Russo VM, Busatto G, Franco M, Rugge M. Helicobacter pylori CagA status, mucosal oxidative damage and gastritis phenotype: a potential pathway to cancer? Helicobacter. 2003;8:227–234.PubMedCrossRefGoogle Scholar
  27. 27.
    Sepulveda AR, Goyal A. Helicobacter pylori and Gastric Neoplasms In: Tan, D. & Lauwers, GY, eds. Advances in Surgical Pathology: Gastric Cancer. Philadelphia, PA: Lippincott Williams and Wilkins; 2011:22–37.Google Scholar
  28. 28.
    Takeda K, Kinoshita I, Shimizu Y, Matsuno Y, Shichinohe T, Dosaka-Akita H. Expression of LGR5, an intestinal stem cell marker, during each stage of colorectal tumorigenesis. Anticancer Res. 2011;31:263–270.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Takeshi Uehara
    • 1
    • 2
  • Deqin Ma
    • 1
  • Yuan Yao
    • 1
  • John P. Lynch
    • 3
  • Knashawn Morales
    • 4
  • Amy Ziober
    • 1
  • Michael Feldman
    • 1
  • Hiroyoshi Ota
    • 2
  • Antonia R. Sepulveda
    • 1
  1. 1.Department of Pathology and Laboratory MedicineUniversity of Pennsylvania School of MedicinePhiladelphiaUSA
  2. 2.Department of Laboratory MedicineShinshu University School of MedicineMatsumotoJapan
  3. 3.Division of Gastroenterology, Department of MedicineUniversity of Pennsylvania School of MedicinePhiladelphiaUSA
  4. 4.Department of BiostatisticsUniversity of Pennsylvania School of MedicinePhiladelphiaUSA

Personalised recommendations