{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,6]],"date-time":"2026-04-06T19:38:31Z","timestamp":1775504311406,"version":"3.50.1"},"reference-count":35,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2020,4,14]],"date-time":"2020-04-14T00:00:00Z","timestamp":1586822400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Pre\u0301mio NOVARTIS | EXCELLENCE in Fundamental Medical Research da NOVA (Pre\u0301mio Novartis | NOVA) 2015","award":["(Pre\u0301mio Novartis | NOVA) 2015"],"award-info":[{"award-number":["(Pre\u0301mio Novartis | NOVA) 2015"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Background: Male breast cancer (BC) is a distinct neoplasm with low but rising incidence, frequently diagnosed as advanced stage disease. Considering the relevance of altered homologous recombination repair (HRR) in male BC, we aimed to explore the biomarker potential of aberrant promoter methylation of ATM, BRCA1, PALB2, RAD51B, and XRCC3. Methods: Formalin-fixed paraffin-embedded (FFPE) tissue samples from 128 male BC patients, paired adjacent normal tissue and 19 gynecomastia cases were collected and assessed by quantitative methylation-specific PCR (qMSP). Non-parametric tests were used to compare methylation levels between tumor and non-tumor samples and to seek for associations with clinicopathological variables. Results: Only RAD51B and XRCC3 disclosed significant differences between tumor and gynecomastia (p &lt; 0.0001 and p = 0.020, respectively). Assembled in a panel, RAD51B and XRCC3 promoter methylation discriminated male BC from gynecomastia with 91.5% sensitivity, 89.5% specificity, and 91.2% accuracy. Moreover, promoter methylation levels were lower in paired non-tumor tissues, comparing to tumor samples. No associations were found between epigenetic alterations and clinicopathological features, as well as with RAD51 and XRCC3 immunoexpression and methylation levels. Conclusion: Quantitative promoter methylation of RAD51B and XRCC3 constitutes a promising and accurate biomarker for male BC. Validation in larger series and in liquid biopsies is warranted to confirm its usefulness in detection and monitoring settings.<\/jats:p>","DOI":"10.3390\/ijms21082715","type":"journal-article","created":{"date-parts":[[2020,4,15]],"date-time":"2020-04-15T04:01:46Z","timestamp":1586923306000},"page":"2715","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Analysis of Epigenetic Alterations in Homologous Recombination DNA Repair Genes in Male Breast Cancer"],"prefix":"10.3390","volume":"21","author":[{"given":"Saudade","family":"Andr\u00e9","sequence":"first","affiliation":[{"name":"Department of Pathology, Portuguese Oncology Institute of Lisboa, 1099-023 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4466-037X","authenticated-orcid":false,"given":"Sandra","family":"P. Nunes","sequence":"additional","affiliation":[{"name":"Cancer Biology & Epigenetics Group\u2014Research Center, Portuguese Oncology Institute of Porto (CI-IPOP), 4200-072 Porto, Portugal"}]},{"given":"Fernanda","family":"Silva","sequence":"additional","affiliation":[{"name":"Medical School, NOVA University, 1169-056 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3171-4666","authenticated-orcid":false,"given":"Rui","family":"Henrique","sequence":"additional","affiliation":[{"name":"Cancer Biology & Epigenetics Group\u2014Research Center, Portuguese Oncology Institute of Porto (CI-IPOP), 4200-072 Porto, Portugal"},{"name":"Department of Pathology, Portuguese Oncology Institute of Porto, 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar\u2013 University of Porto (ICBAS-UP), 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2653-2262","authenticated-orcid":false,"given":"Ana","family":"F\u00e9lix","sequence":"additional","affiliation":[{"name":"Department of Pathology, Portuguese Oncology Institute of Lisboa, 1099-023 Lisboa, Portugal"},{"name":"Medical School, NOVA University, 1169-056 Lisbon, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4186-5345","authenticated-orcid":false,"given":"Carmen","family":"Jer\u00f3nimo","sequence":"additional","affiliation":[{"name":"Cancer Biology & Epigenetics Group\u2014Research Center, Portuguese Oncology Institute of Porto (CI-IPOP), 4200-072 Porto, Portugal"},{"name":"Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar\u2013 University of Porto (ICBAS-UP), 4050-313 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1148\/rg.2018180013","article-title":"Male breast cancer in the age of genetic testing: An opportunity for early detection, tailored therapy, and surveillance","volume":"38","author":"Gao","year":"2018","journal-title":"Radiographics"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1002\/ijc.32106","article-title":"Insight into genetic susceptibility to male breast cancer by multigene panel testing: Results from a multicenter study in Italy","volume":"145","author":"Rizzolo","year":"2019","journal-title":"Int. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1093\/annonc\/mdx651","article-title":"Characterization of male breast cancer: Results of the EORTC 10085\/TBCRC\/BIG\/NABCG International Male Breast Cancer Program","volume":"29","author":"Cardoso","year":"2018","journal-title":"Ann. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Deb, S., Gorringe, K.L., Pang, J.B., Byrne, D.J., Takano, E.A., Investigators, K., Dobrovic, A., and Fox, S.B. (2017). BRCA2 carriers with male breast cancer show elevated tumour methylation. BMC Cancer, 17.","DOI":"10.1186\/s12885-017-3632-7"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Shan, M., Zhang, L., Liu, Y., Gao, C., Kang, W., Yang, W., He, Y., and Zhang, G. (2019). DNA Methylation Profiles and Their Diagnostic Utility in BC. Dis. Markers, 2019.","DOI":"10.1155\/2019\/6328503"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Barros-Silva, D., Marques, C.J., Henrique, R., and Jer\u00f3nimo, C. (2018). Profiling DNA Methylation Based on Next-Generation Sequencing Approaches: New Insights and Clinical Applications. Genes, 9.","DOI":"10.3390\/genes9090429"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1038\/s41431-017-0021-2","article-title":"Contribution of germline deleterious variants in the RAD51 paralogs to breast and ovarian cancers","volume":"25","author":"Golmard","year":"2017","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Heeke, A.L., Pishvaian, M.J., Lynce, F., Xiu, J., Brody, J.R., Chen, W.J., Baker, T.M., Marshall, J.L., and Isaacs, C. Prevalence of Homologous Recombination-Related Gene Mutations Across Multiple Cancer Types. JCO Precis. Oncol., 2018.","DOI":"10.1200\/PO.17.00286"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1830","DOI":"10.1128\/MCB.01521-12","article-title":"ATM- and ATR-mediated phosphorylation of XRCC3 regulates DNA double-strand break-induced checkpoint activation and repair","volume":"33","author":"Somyajit","year":"2013","journal-title":"Mol. Cell Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1002\/cncr.30337","article-title":"Whole-exome sequencing and targeted gene sequencing provide insights into the role of PALB2 as a male breast cancer susceptibility gene","volume":"123","author":"Silvestri","year":"2017","journal-title":"Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Pelttari, L.M., Khan, S., Vuorela, M., Kiiski, J.I., Vilske, S., Nevanlinna, V., Ranta, S., Schleutker, J., Winqvist, R., and Kallioniemi, A. (2016). RAD51B in Familial Breast Cancer. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0153788"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Sullivan, M.R., and Bernstein, K.A. (2018). RAD-ical New Insights into RAD51 Regulation. Genes, 9.","DOI":"10.20944\/preprints201811.0541.v1"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2074","DOI":"10.1002\/cam4.2093","article-title":"Germline mutation landscape of Chinese patients with familial breast\/ovarian cancer in a panel of 22 susceptibility genes","volume":"8","author":"Wang","year":"2019","journal-title":"Cancer Med."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Momozawa, Y., Iwasaki, Y., Hirata, M., Liu, X., Kamatani, Y., Takahashi, A., Sugano, K., Yoshida, T., Murakami, Y., and Matsuda, K. (2019). Germline pathogenic variants in 7636 Japanese patients with prostate cancer and 12,366 controls. J. Natl. Cancer Inst.","DOI":"10.1093\/jnci\/djz124"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"14969","DOI":"10.1007\/s13277-016-5340-3","article-title":"RAD51B as a potential biomarker for early detection and poor prognostic evaluation contributes to tumorigenesis of gastric cancer","volume":"37","author":"Cheng","year":"2016","journal-title":"Tumour Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1007\/s10549-018-4765-3","article-title":"Mutational analysis of candidate genes in Israeli male breast cancer cases","volume":"170","author":"Schayek","year":"2018","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2829","DOI":"10.1002\/cncr.32083","article-title":"Pathogenic and likely pathogenic variants in PALB2, CHEK2, and other known breast cancersusceptibility genes among 1054 BRCA-negative Hispanics with breast cancer","volume":"125","author":"Weitzel","year":"2019","journal-title":"Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"R101","DOI":"10.1186\/bcr3220","article-title":"Promoter hypermethylation in male breast cancer: Analysis by multiplex ligation-dependent probe amplification","volume":"14","author":"Kornegoor","year":"2012","journal-title":"Breast Cancer Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1002\/jcp.24281","article-title":"Different methylation and microRNA expression pattern in male and female familial breast cancer","volume":"228","author":"Pinto","year":"2013","journal-title":"J. Cell Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1565","DOI":"10.1016\/j.molonc.2015.04.013","article-title":"Genome methylation patterns in male breast cancer\u2014Identification of an epitype with hypermethylation of polycomb target genes","volume":"9","author":"Johansson","year":"2015","journal-title":"Mol. Oncol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"19783","DOI":"10.18632\/oncotarget.24856","article-title":"Gene-specific methylation profiles in BRCA-mutation positive and BRCA-mutation negative male breast cancers","volume":"9","author":"Rizzolo","year":"2018","journal-title":"Oncotarget"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.mpdhp.2019.01.004","article-title":"Pathology of the male breast","volume":"25","author":"Shaaban","year":"2019","journal-title":"Diagn. Histopathol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"viii75","DOI":"10.1093\/annonc\/mdt316","article-title":"Male breast cancer: Genetics, epigenetics, and ethical aspects","volume":"24","author":"Rizzolo","year":"2013","journal-title":"Ann. Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1007\/s10549-019-05338-1","article-title":"Mammographic screening in male patients at high risk for breast cancer: Is it worth it?","volume":"177","author":"Marino","year":"2019","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_25","first-page":"644","article-title":"Male breast cancer: Specific biologic characteristics and survival in a Portuguese cohort","volume":"10","author":"Pereira","year":"2019","journal-title":"Mol. Clin. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1007\/s10549-016-4085-4","article-title":"Male breast cancer in a multi-gene panel testing cohort: Insights and unexpected results","volume":"161","author":"Pritzlaff","year":"2017","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_27","first-page":"w13879","article-title":"Impact of family history of breast cancer on tumor characteristics, treatment, risk of second cancer and survival among men with breast cancer","volume":"143","author":"Bouchardy","year":"2013","journal-title":"Swiss Med. Wkly."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.breast.2015.12.001","article-title":"Male breast cancer: Looking for better prognostic subgroups","volume":"26","author":"Abreu","year":"2016","journal-title":"Breast"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1038\/301089a0","article-title":"Hypomethylation distinguishes genes of some human cancers from their normal counterparts","volume":"301","author":"Feinberg","year":"1983","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"75379","DOI":"10.18632\/oncotarget.12211","article-title":"Methylation of RAD51B, XRCC3 and other homologous recombination genes is associated with expression of immune checkpoints and an inflammatory signature in squamous cell carcinoma of the head and neck, lung and cervix","volume":"7","author":"Rieke","year":"2016","journal-title":"Oncotarget"},{"key":"ref_31","first-page":"3","article-title":"Accumulation of genetic and epigenetic alterations in normal cells and cancer risk","volume":"6","author":"Takeshima","year":"2019","journal-title":"NPJ Precis. Oncol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1002\/1097-0142(195309)6:5<963::AID-CNCR2820060515>3.0.CO;2-Q","article-title":"Field cancerization in oral stratified squamous epithelium; clinical implications of multicentric origin","volume":"6","author":"Slaugher","year":"1953","journal-title":"Cancer"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"243","DOI":"10.2217\/epi.12.12","article-title":"DNA methylation and field cancerization","volume":"4","author":"Ramachandran","year":"2012","journal-title":"Epigenomics"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"93","DOI":"10.3322\/caac.21388","article-title":"The Eighth Edition AJCC Cancer Staging Manual: Continuing to Build a Bridge From a Population-Based to a More \"Personalized\" Approach to Cancer Staging","volume":"67","author":"Amin","year":"2017","journal-title":"CA Cancer J. Clin."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1097\/01.ede.0000147512.81966.ba","article-title":"Optimal cut-point and its corresponding Youden Index to discriminate individuals using pooled blood samples","volume":"16","author":"Schisterman","year":"2005","journal-title":"Epidemiology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/8\/2715\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:44:56Z","timestamp":1760363096000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/8\/2715"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,14]]},"references-count":35,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["ijms21082715"],"URL":"https:\/\/doi.org\/10.3390\/ijms21082715","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,4,14]]}}}