Original Article

Epstein-barr virus/Helicobacter pylori coinfection and gastric cancer: the possible role of viral gene expression and shp1 methylation

Abstract

Background and Objectives: Among the various factors involved in the development of gastric cancer (GC), infectious agents are one of the most important causative inducers. This study aimed to investigate the possible role of EBV gene expression on SHP1 methylation in co-infection with Helicobacter pylori in patients with GC.
Materials and Methods: Formalin-fixed paraffin-embedded samples were obtained from 150 patients with gastrointestinal disorders. The presence of the H. pylori and EBV genome were examined by PCR. The expression level of viral gene transcripts and methylation status of the SHP1 cellular gene was assessed by quantitative real-time PCR and methyl-specific PCR.
Results: EBV and H. pylori coinfection were reported in 5.6% of patients. The mean DNA viral load was significant in patients coinfected with cagA-positive H. pylori (P= 0.02). The expression of BZLF1 and EBER was associated with GC. Also, the expression level of BZLF1in GC tissues was significantly higher in coinfection (P = 0.01). SHP1 methylation frequency was higher in the GC group than in the control group (P = 0.04). The correlation between the methylation rate and the H. pylori infection was highly significant (P<0.0001). The strongest positive correlation was observed in GC specimens between SHP1 methylation and H. pylori cagA-positive strains (p= 0.003).
Conclusion: Our results suggested that cagA might involve in the elevation of EBV lytic gene expression and SHP1 methylation, and the development of gastric cancer. Understanding the mechanism of EBV H. pylori - cagA + coinfection, as well as host epigenetic changes, can play an important role in diagnosing and preventing gastric cancer.

1. Balakrishnan M, George R, Sharma A, Graham DY. Changing trends in stomach cancer throughout the world. Curr Gastroenterol Rep 2017; 19: 36.
2. Necula L, Matei L, Dragu D, Neagu AI, Mambet C, Nedeianu S, et al. Recent advances in gastric cancer early diagnosis. World J Gastroenterol 2019; 25: 2029-2044.
3. Ayee R, Ofori MEO, Wright E, Quaye O. Epstein Barr virus associated lymphomas and epithelia cancers in humans. J Cancer 2020; 11: 1737-1750.
4. Shinozaki-Ushiku A, Kunita A, Fukayama M. Update on Epstein-Barr virus and gastric cancer (review). Int J Oncol 2015; 46: 1421-1434.
5. Ishaq S, Nunn L. Helicobacter pylori and gastric cancer: a state of the art review. Gastroenterol Hepatol Bed Bench 2015; 8(Suppl 1): S6-S14.
6. Kashyap D, Baral B, Jakhmola S, Singh AK, Jha HC. Helicobacter pylori and epstein-barr virus coinfection stimulates aggressiveness in gastric cancer through the regulation of gankyrin. mSphere 2021; 6(5): e0075121.
7. Mui UN, Haley CT, Tyring SK. Viral oncology: molecular biology and pathogenesis. J Clin Med 2017; 6: 111.
8. Knight JS, Sharma N, Kalman DE, Robertson ES. A cyclin-binding motif within the amino-terminal homology domain of EBNA3C binds cyclin A and modulates cyclin A-dependent kinase activity in Epstein-Barr virus-infected cells. J Virol 2004; 78: 12857-12867.
9. Ma J, Shen H, Kapesa L, Zeng S. Lauren classification and individualized chemotherapy in gastric cancer. Oncol Lett 2016; 11: 2959-2964.
10. Saju P, Murata-Kamiya N, Hayashi T, Senda Y, Nagase L, Noda S, et al. Host SHP1 phosphatase antagonizes Helicobacter pylori CagA and can be downregulated by Epstein–Barr virus. Nat Microbiol 2016; 1: 16026.
11. Zhou Y, Attygalle AD, Chuang S-S, Diss T, Ye H, Liu H, et al. Angioimmunoblastic T‐cell lymphoma: histological progression associates with EBV and HHV6B viral load. Br J Haematol 2007; 138: 44-53.
12. Baptista M, Cunha JT, Domingues L. DNA-based approaches for dairy products authentication: A review and perspectives. Trends Food Sci Technol 2021; 109: 386-397.
13. Estaji F, Nasr Esfahani BN, Zibaee S, Sanei MH, Moghim S. EPIYA motif genetic characterization from Helicobacter pylori isolates in distinct geographical regions of Iran. Adv Biomed Res 2022; 11: 77.
14. Pandey S, Jha HC, Shukla SK, Shirley MK, Robertson ES. Epigenetic regulation of tumor suppressors by Helicobacter pylori enhances EBV-induced proliferation of gastric epithelial cells. mBio 2018; 9(2): e00649-18.
15. Nakayama A, Abe H, Kunita A, Saito R, Kanda T, Yamashita H, et al. Viral loads correlate with upregulation of PD-L1 and worse patient prognosis in Epstein–Barr Virus-associated gastric carcinoma. PLoS One 2019; 14(1): e0211358.
16. Shukla SK, Prasad KN, Tripathi A, Singh A, Saxena A, Ghoshal UC, et al. Epstein-Barr virus DNA load and its association with Helicobacter pylori infection in gastroduodenal diseases. Braz J Infect Dis 2011; 15: 583-590.
17. Choi H, Lee H, Kim SR, Gho YS, Lee SK. Epstein-Barr virus-encoded microRNA BART15-3p promotes cell apoptosis partially by targeting BRUCE. J Virol 2013; 87: 8135-8144.
18. Pfaffl MW. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res 2001; 29(9): e45.
19. Naseem M, Barzi A, Brezden-Masley C, Puccini A, Berger MD, Tokunaga R, et al. Outlooks on Epstein-Barr virus associated gastric cancer. Cancer Treat Rev 2018; 66: 15-22.
20. Faghihloo E, Saremi MR, Mahabadi M, Akbari H, Saberfar E. Prevalence and characteristics of Epstein barr virus-associated gastric cancer in Iran. Arch Iran Med 2014; 17: 767-770.
21. Leila Z, Arabzadeh SA, Afshar RM, Afshar AA, Mollaei HR. Detection of Epstein-Barr virus and cytomegalovirus in gastric cancers in Kerman, Iran. Asian Pac J Cancer Prev 2016; 17: 2423-2428.
22. Fattahi S, Nikbakhsh N, Taheri H, Ghadami E, Kosari-Monfared M, Amirbozorgi G, et al. Prevalence of multiple infections and the risk of gastric adenocarcinoma development at earlier age. Diagn Microbiol Infect Dis 2018; 92: 62-68.
23. De Souza CRT, Almeida MCA, Khayat AS, Da Silva EL, Soares PC, Chaves LC, et al. Association between Helicobacter pylori, Epstein-Barr virus, human papillomavirus and gastric adenocarcinomas. World J Gastroenterol 2018; 24: 4928-4938.
24. Shukla SK, Prasad KN, Tripathi A, Ghoshal UC, Krishnani N, Husain N. Expression profile of latent and lytic transcripts of epstein–barr virus in patients with gastroduodenal diseases: a study from northern India. J Med Virol 2012; 84: 1289-1297.
25. Ramayanti O, Juwana H, Verkuijlen SAMW, Adham M, Pegtel MD, Greijer AE, et al. Epstein‐Barr virus mRNA profiles and viral DNA methylation status in nasopharyngeal brushings from nasopharyngeal carcinoma patients reflect tumor origin. Int J Cancer 2017; 140: 149-162.
26. Borozan I, Zapatka M, Frappier L, Ferretti V. Analysis of Epstein-Barr virus genomes and expression profiles in gastric adenocarcinoma. J Virol 2018; 92(2): e01239-17.
27. Fekadu S, Kanehiro Y, Kartika AV, Hamada K, Sakurai N, Mizote T, et al. Gastric epithelial attachment of Helicobacter pylori induces EphA2 and NMHC‐IIA receptors for Epstein‐Barr virus. Cancer Sci 2021; 112: 4799-4811.
28. Lee H-H, Chang S-S, Lin S-J, Chua H-H, Tsai T-J, Tsai K, et al. Essential role of PKCδ in histone deacetylase inhibitor-induced Epstein–Barr virus reactivation in nasopharyngeal carcinoma cells. J Gen Virol 2008; 89: 878-883.
29. Brandt S, Wessler S, Hartig R, Backert S. Helicobacter pylori activates protein kinase C delta to control Raf in MAP kinase signalling: role in AGS epithelial cell scattering and elongation. Cell Motil Cytoskeleton 2009; 66: 874-892.
30. Münz C. Latency and lytic replication in Epstein–Barr virus-associated oncogenesis. Nat Rev Microbiol 2019; 17: 691-700.
31. Münz C. Tumor microenvironment conditioning by abortive lytic replication of oncogenic γ-Herpesviruses. Adv Exp Med Biol 2020; 1225: 127-135.
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IssueVol 14 No 6 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v14i6.11265
Keywords
Epstein-barr virus; Helicobacter pylori coinfection; Viral gene expression; SHP1 methylation; Gastric cancer

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How to Cite
1.
Estaji F, Nasr Esfahani B, Zibaee S, Sanei MH, Moghim S. Epstein-barr virus/Helicobacter pylori coinfection and gastric cancer: the possible role of viral gene expression and shp1 methylation. Iran J Microbiol. 2022;14(6):901-912.