{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T14:07:57Z","timestamp":1779199677426,"version":"3.51.4"},"reference-count":26,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2014,12,29]],"date-time":"2014-12-29T00:00:00Z","timestamp":1419811200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Instituto de Salud Carlos III, Fondo Investigaci\u00c3\u00b3n Sanitaria","award":["PI10\/00384; PI13\/00719"],"award-info":[{"award-number":["PI10\/00384; PI13\/00719"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancer"],"published-print":{"date-parts":[[2015,5]]},"abstract":"<jats:sec><jats:title>BACKGROUND<\/jats:title><jats:p>Colorectal cancers (CRCs) that have microsatellite instability (MSI) and mutL homolog 1 (<jats:italic>MLH1<\/jats:italic>) immunoloss are observed in 3 clinical scenarios: Lynch syndrome (LS), sporadic MSI CRC, and Lynch\u2010like syndrome (LLS). v\u2010Raf murine sarcoma viral oncogene homolog B1 (<jats:italic>BRAF<\/jats:italic>) mutational analysis is used to differentiate LS from sporadic MSI CRC. The role of <jats:italic>MLH1<\/jats:italic> promoter methylation status for the differential diagnosis of these clinical forms is not well established. The objectives of this study were: 1) to analyze <jats:italic>MLH1<\/jats:italic> promoter methylation in <jats:italic>MLH1<\/jats:italic>\u2010deficient CRCs by pyrosequencing, and 2) to assess its role in the differential diagnosis of <jats:italic>MLH1<\/jats:italic>\u2010deficient CRCs.<\/jats:p><\/jats:sec><jats:sec><jats:title>METHODS<\/jats:title><jats:p>In total, 165 CRCs were analyzed, including LS (n\u2009=\u200919), MSI <jats:italic>BRAF<\/jats:italic>\u2010mutated CRC (n\u2009=\u200937), MSI <jats:italic>BRAF<\/jats:italic> wild\u2010type CRC (n\u2009=\u200960), and a control group of CRCs without MSI (microsatellite stable [MSS] CRC; n\u2009=\u200949). <jats:italic>MLH1<\/jats:italic> promoter methylation status was analyzed by pyrosequencing, and the ability of different strategies to identify LS was assessed.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>The average \u00b1 standard deviation methylation in LS (9%\u2009\u00b1\u20097%) was significantly lower than that in MSI <jats:italic>BRAF<\/jats:italic>\u2010mutated CRC (42%\u2009\u00b1\u200917%; <jats:italic>P<\/jats:italic>\u2009&lt;\u2009.001) and in MSI <jats:italic>BRAF<\/jats:italic> wild\u2010type CRC (25%\u2009\u00b1\u200919%; <jats:italic>P<\/jats:italic>\u2009=\u2009.002). Somatic <jats:italic>MLH1<\/jats:italic> hypermethylation was detected in 3 patients (15.8%) with LS, in 34 patients (91.9%) with MSI <jats:italic>BRAF<\/jats:italic>\u2010mutated CRC, and in 37 patients (61.7%) with MSI <jats:italic>BRAF<\/jats:italic> wild\u2010type tumors. Patients with MSI <jats:italic>BRAF<\/jats:italic> wild\u2010type, unmethylated tumors (ie, LLS) had a stronger family history of CRC than those who had tumors with <jats:italic>MLH1<\/jats:italic> methylation (<jats:italic>P<\/jats:italic>\u2009&lt;\u2009.05). The sensitivity for ruling out LS was 100% for <jats:italic>BRAF<\/jats:italic> analysis, 84.2% for <jats:italic>MLH1<\/jats:italic> methylation analysis, and 84.2% for the combination of both analyses.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSIONS<\/jats:title><jats:p>Somatic <jats:italic>MLH1<\/jats:italic> promoter methylation occurs in up to 15% of LS CRCs. Somatic <jats:italic>BRAF<\/jats:italic> analysis is the most sensitive strategy for ruling out LS. Patients who have CRCs with loss of MLH1 protein expression and neither <jats:italic>BRAF<\/jats:italic> mutation nor <jats:italic>MLH1<\/jats:italic> methylation resemble patients with LS. <jats:bold><jats:italic>Cancer<\/jats:italic> 2015;121:1395\u20131404.<\/jats:bold> \u00a9 <jats:italic>2014 American Cancer Society<\/jats:italic>.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cncr.29190","type":"journal-article","created":{"date-parts":[[2014,12,29]],"date-time":"2014-12-29T14:01:49Z","timestamp":1419861709000},"page":"1395-1404","source":"Crossref","is-referenced-by-count":52,"title":["Prevalence of somatic mutl homolog 1 promoter hypermethylation in Lynch syndrome colorectal cancer"],"prefix":"10.1002","volume":"121","author":[{"given":"Leticia","family":"Moreira","sequence":"first","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jenifer","family":"Mu\u00f1oz","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M\u00edriam","family":"Cuatrecasas","sequence":"additional","affiliation":[{"name":"Pathology Department Center for Biomedical Diagnostics, Hospital Clinic of Barcelona, University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isabel","family":"Quintanilla","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Liz","family":"Leoz","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sabela","family":"Carballal","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Teresa","family":"Oca\u00f1a","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mar\u00eda","family":"L\u00f3pez\u2010Cer\u00f3n","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria","family":"Pellise","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergi","family":"Castellv\u00ed\u2010Bel","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rodrigo","family":"Jover","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology University General Hospital of Alicante Alicante Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Montserrat","family":"Andreu","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology Hospital del Mar Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angel","family":"Carracedo","sequence":"additional","affiliation":[{"name":"Genomic Medicine Group, Galician Public Foundation for Genomic Medicine Center for Biomedical Network Research on Rare Diseases University of Santiago de Compostela Galicia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosa Maria","family":"Xicola","sequence":"additional","affiliation":[{"name":"Department of Digestive Diseases and Nutrition University of Illinois at Chicago Chicago Illinois"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xavier","family":"Llor","sequence":"additional","affiliation":[{"name":"Department of Digestive Diseases and Nutrition University of Illinois at Chicago Chicago Illinois"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clement Richard","family":"Boland","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Division of Gastroenterology Charles A. Sammons Cancer Center and Baylor Research Institute Baylor University Medical Center Dallas Texas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ajay","family":"Goel","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Division of Gastroenterology Charles A. Sammons Cancer Center and Baylor Research Institute Baylor University Medical Center Dallas Texas"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antoni","family":"Castells","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesc","family":"Balaguer","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Barcelona Hospital Clinic Networked Biomedical Research Center on Hepatic and Digestive Diseases (CIBERehd) August Pi i Sunyer Biomedical Research Institute (IDIBAPS) University of Barcelona Barcelona Catalonia Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2014,12,29]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.293.16.2028"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.ejhg.5201584"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/363558a0"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmoldx.2010.12.004"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2005.11.032"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2012-304356"},{"key":"e_1_2_8_8_1","first-page":"3455","article-title":"Hypermethylation of the hMLH1 promoter in colon cancer with microsatellite instability","volume":"58","author":"Cunningham JM","year":"1998","journal-title":"Cancer Res"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2013.01.044"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2013.12.002"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00428-013-1470-9"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.22544"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.2353\/jmoldx.2010.090212"},{"key":"e_1_2_8_14_1","first-page":"2029","article-title":"Methylation of CpG in a small region of the hMLH1 promoter invariably correlates with the absence of gene expression","volume":"59","author":"Deng G","year":"1999","journal-title":"Cancer Res"},{"key":"e_1_2_8_15_1","first-page":"721","article-title":"Pyrosequencing assays to study promoter CpG site methylation of the O6\u2010MGMT, hMLH1, p14ARF, p16INK4a, RASSF1A, and APC1A genes","volume":"21","author":"Lof\u2010Ohlin ZM","year":"2009","journal-title":"Oncol Rep"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2011-300041"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-10-0622"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng1342"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.2012.13088"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cgh.2007.10.011"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrclinonc.2010.18"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2407-10-227"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1136\/jmedgenet-2011-100714"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-06-3064"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/ejhg.2011.277"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)65048-1"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa064522"}],"container-title":["Cancer"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcncr.29190","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fcncr.29190","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/acsjournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cncr.29190","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T18:18:58Z","timestamp":1760293138000},"score":1,"resource":{"primary":{"URL":"https:\/\/acsjournals.onlinelibrary.wiley.com\/doi\/10.1002\/cncr.29190"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,12,29]]},"references-count":26,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2015,5]]}},"alternative-id":["10.1002\/cncr.29190"],"URL":"https:\/\/doi.org\/10.1002\/cncr.29190","archive":["Portico"],"relation":{},"ISSN":["0008-543X","1097-0142"],"issn-type":[{"value":"0008-543X","type":"print"},{"value":"1097-0142","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,12,29]]}}}