The EPIYA-ABCC motif pattern in CagA of Helicobacter pylori is associated with peptic ulcer and gastric cancer in Mexican population

BMC Gastroenterology

Gloria Fernández-Tilapa³  Berenice Illades-Aguiar²  Oscar del Moral-Hernández²  Miguel Ángel Rodríguez²  Oscar Peralta-Zaragoza¹  José de Sampedro⁴  Salomón Reyes⁵  Adolfo Román-Román³  Tomás Manuel Poblete³  Fredy Omar Beltrán-Anaya³ 

[1]Department of Chronic Infections and Cancer, Infectious Diseases Research Center, National Institute of Public Health, Av. Universidad No. 655, Cerrada los Pinos y Caminera, Colonia Santa María Ahuacatitlan, Cuernavaca C.P. 62100, Morelos, Mexico

[2]Laboratory of Molecular Biomedicine, Academic Unit of Chemical-Biological Sciences, Autonomous University of Guerrero, Chilpancingo C.P. 39090, Guerrero, Mexico

[3]Clinical Research Laboratory, Academic Unit of Chemical-Biological Sciences, Autonomous University of Guerrero, Chilpancingo C.P. 39090, Guerrero, Mexico

[4]General Hospital “Dr. Raymundo Abarca Alarcón”, Chilpancingo C.P. 39090, Guerrero, Mexico

[5]State Institute of Oncology “Dr. Arturo Beltrán Ortega”, Acapulco C. P. 39570, Guerrero, Mexico

关键词: H. pylori;    CagA;    EPIYA;    CRPIA;    cagA gene 3′ region;   

Others  :  1121722
DOI  :  10.1186/s12876-014-0223-9

 received in 2014-08-13, accepted in 2014-12-17,  发布年份 2014

【 摘 要 】

Background

Helicobacter pylori chronic infection is associated with chronic gastritis, peptic ulcer, and gastric cancer. Cytotoxin-associated gene A (cagA)-positive H. pylori strains increase the risk of gastric pathology. The carcinogenic potential of CagA is linked to its polymorphic EPIYA motif variants. The goals of this study were to investigate the frequency of cagA-positive Helicobacter pylori in Mexican patients with gastric pathologies and to assess the association of cagA EPIYA motif patterns with peptic ulcer and gastric cancer.

Methods

A total of 499 patients were studied; of these, 402 had chronic gastritis, 77 had peptic ulcer, and 20 had gastric cancer. H. pylori DNA, cagA, and the EPIYA motifs were detected in total DNA from gastric biopsies by PCR. The type and number of EPIYA segments were determined by the electrophoretic patterns. To confirm the PCR results, 20 amplicons of the cagA 3′ variable region were sequenced, and analyzed in silico, and the amino acid sequence was predicted with MEGA software, version 5. The odds ratio (OR) was calculated to determine the associations between the EPIYA motif type and gastric pathology and between the number of EPIYA-C segments and peptic ulcers and gastric cancer.

Results

H. pylori DNA was found in 287 (57.5%) of the 499 patients, and 214 (74%) of these patients were cagA-positive. The frequency of cagA-positive H. pylori was 74.6% (164/220) in chronic gastritis patients, 73.6% (39/53) in peptic ulcer patients, and 78.6% (11/14) in gastric cancer patients. The EPIYA-ABC pattern was more frequently observed in chronic gastritis patients (79.3%, 130/164), while the EPIYA-ABCC sequence was more frequently observed in peptic ulcer (64.1%, 25/39) and gastric cancer patients (54.5%, 6/11). However, the risks of peptic ulcer (OR = 7.0, 95% CI = 3.3–15.1; p < 0.001) and gastric cancer (OR = 5.9, 95% CI = 1.5–22.1) were significantly increased in individuals who harbored the EPIYA-ABCC cagA gene pattern.

Conclusions

cagA-positive H. pylori is highly prevalent in southern Mexico, and all CagA variants were of the western type. The cagA alleles that code for EPIYA-ABCC motif patterns are associated with peptic ulcers and gastric cancer.

【 授权许可】

   
2014 Beltrán-Anaya et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]de Martel C, Forman D, Plummer M: Gastric cancer: epidemiology and risk factors. Gastroenterol Clin N Am 2013, 42:219-40.
• [2]Wroblewski LE, Peek RM Jr, Wilson KT: Helicobacter pylori and gastric cancer: factors that modulate disease risk. Clin Microbiol Rev 2010, 23:713-39.
• [3]Hatakeyama M: Helicobacter pylori and gastric carcinogenesis. J Gastroenterol 2009, 44:239-48.
• [4]Kuipers EJ, Perez-Perez GI, Meuwissen SG, Blaser MJ: Helicobacter pylori and atrophic gastritis: importance of the cagA status. J Natl Cancer Inst 1995, 87:1777-80.
• [5]Proenca Modena JL, Lopes Sales AI, Olszanski Acrani G, Russo R, Vilela Ribeiro MA, Fukuhara Y, da Silveira WD, Modena JL, de Oliveira RB, Brocchi M: Association between Helicobacter pylori genotypes and gastric disorders in relation to the cag pathogenicity island. Diagn Microbiol Infect Dis 2007, 59:7-16.
• [6]Secka O, Antonio M, Berg DE, Tapgun M, Bottomley C, Thomas V, Walton R, Corrah T, Thomas JE, Adegbola RA: Mixed infection with cagA positive and cagA negative strains of Helicobacter pylori lowers disease burden in The Gambia. PLoS One 2011, 6:e27954.
• [7]Basso D, Zambon CF, Letley DP, Stranges A, Marchet A, Rhead JL, Schiavon S, Guariso G, Ceroti M, Nitti D, Rugge M, Plebani M, Atherton JC: Clinical relevance of Helicobacter pylori cagA and vacA gene polymorphisms. Gastroenterology 2008, 135:91-9.
• [8]Camorlinga-Ponce M, Flores-Luna L, Lazcano-Ponce E, Herrero R, Bernal-Sahagun F, Abdo-Francis JM, Aguirre-Garcia J, Munoz N, Torres J: Age and severity of mucosal lesions influence the performance of serologic markers in Helicobacter pylori-associated gastroduodenal pathologies. Cancer Epidemiol Biomarkers Prev 2008, 17:2498-504.
• [9]Figura N, Vindigni C, Covacci A, Presenti L, Burronim D, Vernillo R, Banducci T, Roviello F, Marrelli D, Biscontri M, Kristodhullu S, Gennari C, Vaira D: cagA positive and negative Helicobacter pylori strains are simultaneously present in the stomach of most patients with non-ulcer dyspepsia: relevance to histological damage. Gut 1998, 42:772-8.
• [10]Flores-Luna L, Camorlinga-Ponce M, Hernandez-Suarez G, Kasamatsu E, Martinez ME, Murillo R, Lazcano E, Torres J: The utility of serologic tests as biomarkers for Helicobacter pylori-associated precancerous lesions and gastric cancer varies between Latin American countries. Cancer Causes Control 2013, 24:241-8.
• [11]Huang JQ, Zheng GF, Sumanac K, Irvine EJ, Hunt RH: Meta-analysis of the relationship between cagA seropositivity and gastric cancer. Gastroenterology 2003, 125:1636-44.
• [12]Palli D, Masala G, Del Giudice G, Plebani M, Basso D, Berti D, Numans ME, Ceroti M, Peeters PH, de Mesquita HB B, Buchner FL, Clavel-Chapelon F, Boutron-Ruault MC, Krogh V, Saieva C, Vineis P, Panico S, Tumino R, Nyrén O, Simán H, Berglund G, Hallmans G, Sanchez MJ, Larrãnaga N, Barricarte A, Navarro C, Quiros JR, Key T, Allen N, Bingham S: CagA+ Helicobacter pylori infection and gastric cancer risk in the EPIC-EURGAST study. Int J Cancer 2007, 120(4):859-67.
• [13]Sahara S, Sugimoto M, Vilaichone RK, Mahachai V, Miyajima H, Furuta T, Yamaoka Y: Role of Helicobacter pylori cagA EPIYA motif and vacA genotypes for the development of gastrointestinal diseases in Southeast Asian countries: a meta-analysis. BMC Infect Dis 2012, 12:223. BioMed Central Full Text
• [14]Satomi S, Yamakawa A, Matsunaga S, Masaki R, Inagaki T, Okuda T, Suto H, Ito Y, Yamazaki Y, Kuriyama M, Keida Y, Kutsumi H, Azuma T: Relationship between the diversity of the cagA gene of Helicobacter pylori and gastric cancer in Okinawa, Japan. J Gastroenterol 2006, 41:668-73.
• [15]Sicinschi LA, Correa P, Peek RM, Camargo MC, Piazuelo MB, Romero-Gallo J, Hobbs SS, Krishna U, Delgado A, Mera R, Bravo LE, Schneider BG: CagA C-terminal variations in Helicobacter pylori strains from Colombian patients with gastric precancerous lesions. Clin Microbiol Infect 2010, 16:369-78.
• [16]Matteo MJ, Granados G, Perez CV, Olmos M, Sanchez C, Catalano M: Helicobacter pylori cag pathogenicity island genotype diversity within the gastric niche of a single host. J Med Microbiol 2007, 56(Pt 5):664-9.
• [17]Nomura AM, Lee J, Stemmermann GN, Nomura RY, Perez-Perez GI, Blaser MJ: Helicobacter pylori CagA seropositivity and gastric carcinoma risk in a Japanese American population. J Infect Dis 2002, 186:1138-44.
• [18]Backert S, Ziska E, Brinkmann V, Zimny-Arndt U, Fauconnier A, Jungblut PR, Naumann M, Meyer TF: Translocation of the Helicobacter pylori CagA protein in gastric epithelial cells by a type IV secretion apparatus. Cell Microbiol 2000, 2:155-64.
• [19]Rohde M, Puls J, Buhrdorf R, Fischer W, Haas R: A novel sheathed surface organelle of the Helicobacter pylori cag type IV secretion system. Mol Microbiol 2003, 49:219-34.
• [20]Mueller D, Tegtmeyer N, Brandt S, Yamaoka Y, De Poire E, Sgouras D, Wessler S, Torres J, Smolka A, Backert S: c-Src and c-Abl kinases control hierarchic phosphorylation and function of the CagA effector protein in Western and East Asian Helicobacter pylori strains. J Clin Invest 2012, 122:1553-66.
• [21]Selbach M, Moese S, Hauck CR, Meyer TF, Backert S: Src is the kinase of the Helicobacter pylori CagA protein in vitro and in vivo. J Biol Chem 2002, 277:6775-8.
• [22]Tegtmeyer N, Backert S: Role of Abl and Src family kinases in actin-cytoskeletal rearrangements induced by the Helicobacter pylori CagA protein. Eur J Cell Biol 2011, 90:880-90.
• [23]Murata-Kamiya N: Pathophysiological functions of the CagA oncoprotein during infection by Helicobacter pylori. Microbes Infect 2011, 13:799-807.
• [24]Backert S, Tegtmeyer N, Selbach M: The versatility of Helicobacter pylori CagA effector protein functions: the master key hypothesis. Helicobacter 2010, 15:163-76.
• [25]Higashi H, Tsutsumi R, Muto S, Sugiyama T, Azuma T, Asaka M, Hatakeyama M: SHP-2 tyrosine phosphatase as an intracellular target of Helicobacter pylori CagA protein. Science 2002, 295:683-6.
• [26]Suzuki M, Kiga K, Kersulyte D, Cok J, Hooper CC, Mimuro H, Sanada T, Suzuki S, Oyama M, Kozuka-Hata H, Kamiya S, Zou QM, Gilman RH, Berg DE, Sasakawa C: Attenuated CagA oncoprotein in Helicobacter pylori from Amerindians in Peruvian Amazon. J Biol Chem 2011, 286:29964-72.
• [27]Suzuki M, Mimuro H, Kiga K, Fukumatsu M, Ishijima N, Morikawa H, Nagai S, Koyasu S, Gilman RH, Kersulyte D, Berg DE, Sasakawa C: Helicobacter pylori CagA phosphorylation-independent function in epithelial proliferation and inflammation. Cell Host Microbe 2009, 5:23-34.
• [28]Yamazaki S, Yamakawa A, Ito Y, Ohtani M, Higashi H, Hatakeyama M, Azuma T: The CagA protein of Helicobacter pylori is translocated into epithelial cells and binds to SHP-2 in human gastric mucosa. J Infect Dis 2003, 187:334-7.
• [29]Quiroga AJ, Huertas A, Combita AL, Bravo MM: Variation in the number of EPIYA-C repeats in CagA protein from Colombian Helicobacter pylori strains and its ability middle to induce hummingbird phenotype in gastric epithelial cells. Biomedica 2010, 30:251-8.
• [30]Salih BA, Bolek BK, Arikan S: DNA sequence analysis of cagA 3′ motifs of Helicobacter pylori strains from patients with peptic ulcer diseases. J Med Microbiol 2010, 59(Pt 2):144-8.
• [31]Camorlinga-Ponce M, Perez-Perez G, Gonzalez-Valencia G, Mendoza I, Penaloza-Espinosa R, Ramos I, Kersulyte D, Reyes-Leon A, Romo C, Granados J, Muñoz L, Berg DE, Torres J: Helicobacter pylori genotyping from American indigenous groups shows novel Amerindian vacA and cagA alleles and Asian. African and European admixture. PLoS One 2011, 6:e27212.
• [32]Bosques-Padilla FJ, Tijerina-Menchaca R, Perez-Perez GI, Flores-Gutierrez JP, Garza-Gonzalez E: Comparison of Helicobacter pylori prevalence in symptomatic patients in northeastern Mexico with the rest of the country: its association with gastrointestinal disease. Arch Med Res 2003, 34:60-3.
• [33]Lopez-Carrillo L, Camargo MC, Schneider BG, Sicinschi LA, Hernandez-Ramirez RU, Correa P, Cebrian ME: Capsaicin consumption, Helicobacter pylori CagA status and IL1B-31C > T genotypes: a host and environment interaction in gastric cancer. Food Chem Toxicol 2012, 50:2118-22.
• [34]Lopez-Carrillo L, Torres-Lopez J, Galvan-Portillo M, Munoz L, Lopez-Cervantes M: Helicobacter pylori-CagA seropositivity and nitrite and ascorbic acid food intake as predictors for gastric cancer. Eur J Cancer 2004, 40:1752-9.
• [35]Sicinschi LA, Lopez-Carrillo L, Camargo MC, Correa P, Sierra RA, Henry RR, Chen J, Zabaleta J, Piazuelo MB, Schneider BG: Gastric cancer risk in a Mexican population: role of Helicobacter pylori CagA positive infection and polymorphisms in interleukin-1 and −10 genes. Int J Cancer 2006, 118:649-57.
• [36]Torres J, Lopez L, Lazcano E, Camorlinga M, Flores L, Munoz O: Trends in Helicobacter pylori infection and gastric cancer in Mexico. Cancer Epidemiol Biomarkers Prev 2005, 14:1874-7.
• [37]Reyes-Leon A, Atherton JC, Argent RH, Puente JL, Torres J: Heterogeneity in the activity of Mexican Helicobacter pylori strains in gastric epithelial cells and its association with diversity in the cagA gene. Infect Immun 2007, 75:3445-54.
• [38]Rizzato C, Torres J, Plummer M, Munoz N, Franceschi S, Camorlinga-Ponce M, Fuentes-Panana EM, Canzian F, Kato I: Variations in Helicobacter pylori cytotoxin-associated genes and their influence in progression to gastric cancer: implications for prevention. PLoS One 2012, 7:e29605.
• [39]Green MR, Sambrook J: Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York; 2012.
• [40]Roman-Roman A, Giono-Cerezo S, Camorlinga-Ponce M, Martinez-Carrillo DN, Loaiza-Loeza S, Fernandez-Tilapa G: vacA genotypes of Helicobacter pylori in the oral cavity and stomach of patients with chronic gastritis and gastric ulcer. Enferm Infecc Microbiol Clin 2013, 31:130-5.
• [41]Argent RH, Zhang Y, Atherton JC: Simple method for determination of the number of Helicobacter pylori CagA variable-region EPIYA tyrosine phosphorylation motifs by PCR. J Clin Microbiol 2005, 43:791-5.
• [42]Schmidt HM, Goh KL, Fock KM, Hilmi I, Dhamodaran S, Forman D, Mitchell H: Distinct cagA EPIYA motifs are associated with ethnic diversity in Malaysia and Singapore. Helicobacter 2009, 14:256-63.
• [43]Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S: MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011, 28:2731-9.
• [44]Queiroz DM, Silva CI, Goncalves MH, Braga-Neto MB, Fialho AB, Fialho AM, Rocha GA, Rocha AM, Batista SA, Guerrant RL, Lima AA, Braga LL: Higher frequency of cagA EPIYA-C phosphorylation sites in H. pylori strains from first-degree relatives of gastric cancer patients. BMC Gastroenterol 2012, 12:107. BioMed Central Full Text
• [45]Paniagua GL, Monroy E, Rodriguez R, Arroniz S, Rodriguez C, Cortes JL, Camacho A, Negrete E, Vaca S: Frequency of vacA, cagA and babA2 virulence markers in Helicobacter pylori strains isolated from Mexican patients with chronic gastritis. Ann Clin Microbiol Antimicrob 2009, 8:14. BioMed Central Full Text
• [46]Azuma T, Ohtani M, Yamazaki Y, Higashi H, Hatakeyama M: Meta-analysis of the relationship between CagA seropositivity and gastric cancer. Gastroenterology 2004, 126:1926-7. author reply 1927–1928
• [47]Kumar S, Kumar A, Dixit VK: Diversity in the cag pathogenicity island of Helicobacter pylori isolates in populations from North and South India. J Med Microbiol 2010, 59(Pt 1):32-40.
• [48]Batista SA, Rocha GA, Rocha AM, Saraiva IE, Cabral MM, Oliveira RC, Queiroz DM: Higher number of Helicobacter pylori CagA EPIYA C phosphorylation sites increases the risk of gastric cancer, but not duodenal ulcer. BMC Microbiol 2011, 11:61. BioMed Central Full Text
• [49]Acosta N, Quiroga A, Delgado P, Bravo MM, Jaramillo C: Helicobacter pylori CagA protein polymorphisms and their lack of association with pathogenesis. World J Gastroenterol 2010, 16:3936-43.
• [50]Torres LE, Gonzalez L, Melian K, Alonso J, Moreno A, Hernandez M, Reyes O, Bermudez L, Campos J, Perez-Perez G, Rodríguez BL: EPIYA motif patterns among Cuban Helicobacter pylori CagA positive strains. Biomedica 2012, 32:23-31.
• [51]Kersulyte D, Kalia A, Gilman RH, Mendez M, Herrera P, Cabrera L, Velapatino B, Balqui J, Paredes Puente dela Vega F, Rodriguez Ulloa CA, Cok J, Hooper CC, Dailide G, Tamma S, Berg DE: Helicobacter pylori from Peruvian amerindians: traces of human migrations in strains from remote Amazon, and genome sequence of an Amerind strain. PLoS One 2010, 5:e15076.
• [52]Sgouras DN, Panayotopoulou EG, Papadakos K, Martinez-Gonzalez B, Roumbani A, Panayiotou J, VanVliet-Constantinidou C, Mentis AF, Roma-Giannikou E: CagA and VacA polymorphisms do not correlate with severity of histopathological lesions in Helicobacter pylori-infected Greek children. J Clin Microbiol 2009, 47:2426-34.
• [53]Furuta Y, Yahara K, Hatakeyama M, Kobayashi I: Evolution of cagA oncogene of Helicobacter pylori through recombination. PLoS One 2011, 6:e23499.
• [54]Ishikawa S, Ohta T, Hatakeyama M: Stability of Helicobacter pylori CagA oncoprotein in human gastric epithelial cells. FEBS Lett 2009, 583:2414-8.
• [55]Ferlay J, Shin H, Bray F, Forman D, Mathers C, Parkin D: GLOBOCAN 2008 v2.0, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10 [Internet]. Lyon, France: International Agency for Research on Cancer; 2010. Available from: http://globocan.iarc.fr.
• [56]Tarkhashvili N, Chakvetadze N, Mebonia N, Chubinidze M, Bakanidze L, Shengelidze V, Mirtskhulava M, Chachava T, Katsitadze G, Gabunia U, Kordzaia D, Imnadze P, Guarner J, Sobel J: Traditional risk factors for Helicobacter pylori infection not found among patients undergoing diagnostic upper endoscopy-Republic of Georgia, 2007–2008. Int J Infect Dis 2012, 16:e697-702.
• [57]Wu Y, Fan Y, Jiang Y, Wang Y, Liu H, Wei M: Analysis of risk factors associated with precancerous lesion of gastric cancer in patients from eastern China: a comparative study. J Cancer Res Ther 2013, 9:205-9.