{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T19:04:00Z","timestamp":1787166240732,"version":"build-2736575974"},"reference-count":56,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,1,20]],"date-time":"2021-01-20T00:00:00Z","timestamp":1611100800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Oncol."],"abstract":"<jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>Determine the presence and prognostic value of human papillomavirus (HPV), Epstein-Barr virus (EBV), Merkel cell polyomavirus (MCPyV), and cell cycle proteins in head and neck squamous cell carcinoma (HNSCC) of non-smokers and non-drinkers (NSND).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Clinical characteristics and tumors of 119 NSND with HNSCC were retrospectively collected and analyzed on tissue microarrays. RNAscope\n                      <jats:italic>in situ<\/jats:italic>\n                      hybridization (ISH) was used to screen for the presence of HPV and MCPyV mRNA. Immunohistochemistry was performed for expression of p16 as surrogate marker for HPV, Large T-antigen for MCPyV, and cell cycle proteins p53 and pRb. Positive virus results were confirmed with polymerase chain reaction. For EBV, EBV encoded RNA ISH was performed. Differences in 5-year survival between virus positive and negative tumors were determined by log rank analysis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>All oropharyngeal tumors (OPSCC) (n = 10) were HPV-positive, in addition to one oral (OSCC) and one nasopharyngeal tumor (NPSCC). The other three NPSCC were EBV-positive. MCPyV was not detected. Patients with HPV or EBV positive tumors did not have a significantly better 5-year disease free or overall survival. Over 70% of virus negative OSCC showed mutant-type p53 expression.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>In this cohort, all OPSCC and NPSCC showed HPV or EBV presence. Besides one OSCC, all other oral (n = 94), hypopharyngeal (n = 1), and laryngeal (n = 9) tumors were HPV, EBV, and MCPyV negative. This argues against a central role of these viruses in the ethiopathogenesis of tumors outside the oro- and nasopharynx in NSND. So, for the majority of NSND with virus negative OSCC, more research is needed to understand the carcinogenic mechanisms in order to consider targeted therapeutic options.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fonc.2020.560434","type":"journal-article","created":{"date-parts":[[2021,1,20]],"date-time":"2021-01-20T01:56:57Z","timestamp":1611107817000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":20,"title":["Presence of Human Papillomavirus and Epstein\u2013Barr Virus, but Absence of Merkel Cell Polyomavirus, in Head and Neck Cancer of Non-Smokers and Non-Drinkers"],"prefix":"10.3389","volume":"10","author":[{"given":"Frans J.","family":"Mulder","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faisal","family":"Klufah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Famke M. E.","family":"Janssen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farzaneh","family":"Farshadpour","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan M.","family":"Willems","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Remco","family":"de Bree","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Axel","family":"zur Hausen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mari F. C. M.","family":"van den Hout","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernd","family":"Kremer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ernst-Jan M.","family":"Speel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,1,20]]},"reference":[{"key":"B1","volume-title":"TNM Classification of Malignant Tumours 8th Edition","author":"Brierley","year":"2016"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/j.oraloncology.2012.06.019","article-title":"Human papillomavirus related head and neck cancer survival: a systematic review and meta-analysis","volume":"48","author":"O\u2019Rorke","year":"2012","journal-title":"Oral Oncol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.22851","article-title":"Survival of squamous cell carcinoma of the head and neck in relation to human papillomavirus infection: review and meta-analysis","volume":"121","author":"Ragin","year":"2007","journal-title":"Int J Cancer"},{"key":"B4","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1016\/j.ctro.2017.10.005","article-title":"Treatment de-escalation for HPV-driven oropharyngeal cancer: Where do we stand","volume":"8","author":"Mirghani","year":"2018","journal-title":"Clin Trans Radiat Oncol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"djv403","DOI":"10.1093\/jnci\/djv403","article-title":"HPV Involvement in Head and Neck Cancers: Comprehensive Assessment of Biomarkers in 3680 Patients","volume":"108","author":"Castellsague","year":"2016","journal-title":"J Natl Cancer Institute"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1002\/hed.23942","article-title":"Rising prevalence of human papillomavirus-related oropharyngeal cancer in Australia over the last 2 decades","volume":"38","author":"Hong","year":"2016","journal-title":"Head Neck"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.24911","article-title":"Biological and clinical relevance of transcriptionally active human papillomavirus (HPV) infection in oropharynx squamous cell carcinoma","volume":"126","author":"Jung","year":"2010","journal-title":"Int J Cancer"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1016\/j.oraloncology.2018.01.010","article-title":"Double positivity for HPV-DNA\/p16ink4a is the biomarker with strongest diagnostic accuracy and prognostic value for human papillomavirus related oropharyngeal cancer patients","volume":"78","author":"Mena","year":"2018","journal-title":"Oral Oncol"},{"key":"B9","doi-asserted-by":"publisher","first-page":"S68","DOI":"10.1038\/modpathol.2016.189","article-title":"Virus-associated neoplasms of the nasopharynx and sinonasal tract: diagnostic problems","volume":"30","author":"Chan","year":"2017","journal-title":"Modern Pathol"},{"key":"B10","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1016\/j.ctrv.2018.02.008","article-title":"Immunotherapy in head and neck cancers: A new challenge for immunologists, pathologists and clinicians","volume":"65","author":"Outh-Gauer","year":"2018","journal-title":"Cancer Treat Rev"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1002\/hed.25131","article-title":"Presence of lytic Epstein-Barr virus infection in nasopharyngeal carcinoma","volume":"40","author":"Yu","year":"2018","journal-title":"Head Neck"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1126\/science.1152586","article-title":"Clonal integration of a polyomavirus in human Merkel cell carcinoma","volume":"319","author":"Feng","year":"2008","journal-title":"Sci (N Y NY)"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-08-0949","article-title":"Frequent detection of Merkel cell polyomavirus in human Merkel cell carcinomas and identification of a unique deletion in the VP1 gene","volume":"68","author":"Kassem","year":"2008","journal-title":"Cancer Res"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3892\/mco.2015.629","article-title":"Low prevalence of Merkel cell polyomavirus with low viral loads in oral and maxillofacial tumours or tumour-like lesions from immunocompetent patients: Absence of Merkel cell polyomavirus-associated neoplasms","volume":"3","author":"Tanio","year":"2015","journal-title":"Mol Clin Oncol"},{"key":"B15","doi-asserted-by":"publisher","first-page":"695","DOI":"10.1002\/jmv.24385","article-title":"Detection of human polyomaviruses MCPyV, HPyV6, and HPyV7 in malignant and non-malignant tonsillar tissues","volume":"88","author":"Salakova","year":"2016","journal-title":"J Med Virol"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.oraloncology.2017.09.021","article-title":"A pilot study of Merkel cell polyomavirus in squamous cell carcinoma of the tongue","volume":"74","author":"Hamiter","year":"2017","journal-title":"Oral Oncol"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.24953","article-title":"Low viral load of Merkel cell polyomavirus in Iranian patients with head and neck squamous cell carcinoma: Is it clinically important","volume":"90","author":"Mohebbi","year":"2018","journal-title":"J Med Virol"},{"key":"B18","article-title":"Frequency and role of HPV in the progression of epithelial dysplasia to oral cancer","volume":"30","author":"Angiero","year":"2010","journal-title":"Anticancer Res"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1093\/abbs\/gmp025","article-title":"Identification of aberrant cell cycle regulation in Epstein-Barr virus-associated nasopharyngeal carcinoma by cDNA microarray and gene set enrichment analysis","volume":"41","author":"Zhang","year":"2009","journal-title":"Acta Biochim Biophys Sin (Shanghai)"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1038\/s41571-018-0103-2","article-title":"The biology and treatment of Merkel cell carcinoma: current understanding and research priorities","volume":"15","author":"Harms","year":"2018","journal-title":"Nat Rev Clin Oncol"},{"key":"B21","doi-asserted-by":"publisher","first-page":"213","DOI":"10.3390\/cancers10070213","article-title":"Human Oncoviruses and p53 Tumor Suppressor Pathway Deregulation at the Origin of Human Cancers","volume":"10","author":"Tornesello","year":"2018","journal-title":"Cancers"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1016\/j.oraloncology.2008.07.008","article-title":"Oropharyngeal carcinoma in non-smokers and non-drinkers: a role for HPV","volume":"45","author":"Andrews","year":"2009","journal-title":"Oral Oncol"},{"key":"B23","doi-asserted-by":"publisher","first-page":"806345","DOI":"10.4061\/2011\/806345","article-title":"Human Papillomavirus and Oropharyngeal Squamous Cell Carcinoma: A Case-Control Study regarding Tobacco and Alcohol Consumption","volume":"2011","author":"Farshadpour","year":"2011","journal-title":"Pathol Res Int"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1007\/s00428-010-1037-y","article-title":"The role of high-risk human papillomavirus infection in oral and oropharyngeal squamous cell carcinoma in non-smoking and non-drinking patients: a clinicopathological and molecular study of 46 cases","volume":"458","author":"Laco","year":"2011","journal-title":"Virchows Archiv An Int J Pathol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1002\/hed.26513","article-title":"Evidence for different molecular parameters in head and neck squamous cell carcinoma of nonsmokers and nondrinkers: Systematic review and meta-analysis on HPV, p16, and TP53","volume":"41","author":"Mulder","year":"2020","journal-title":"Head Neck"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0714.2008.00657.x","article-title":"Head and neck squamous cell carcinoma in non-smoking and non-drinking patients with multiple tumors: etiologic significance of p53 and Ki-67 in non-tumorous epithelium","volume":"37","author":"Farshadpour","year":"2008","journal-title":"J Oral Pathol Med Off Publ Int Assoc Oral Pathol Am Acad Oral Pathol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1016\/j.neo.2018.10.003","article-title":"Neuroendocrine Key Regulator Gene Expression in Merkel Cell Carcinoma","volume":"20","author":"Chteinberg","year":"2018","journal-title":"Neoplasia"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1002\/ijc.28534","article-title":"P16(INK4A) immunostaining is a strong indicator for high-risk-HPV-associated oropharyngeal carcinomas and dysplasias, but is unreliable to predict low-risk-HPV-infection in head and neck papillomas and laryngeal dysplasias","volume":"134","author":"Mooren","year":"2014","journal-title":"Int J Cancer"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.3892\/ol.2015.3651","article-title":"Expression of mutant p53 in oral squamous cell carcinoma is correlated with the effectiveness of intra-arterial chemotherapy","volume":"10","author":"Li","year":"2015","journal-title":"Oncol Lett"},{"key":"B30","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1186\/s40463-018-0298-3","article-title":"Prognostic significance of cell cycle-associated proteins p16, pRB, cyclin D1 and p53 in resected oropharyngeal carcinoma","volume":"47","author":"Plath","year":"2018","journal-title":"J Otolaryngol Head Neck Surg"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1016\/j.humpath.2014.12.001","article-title":"Merkel cell carcinoma: histopathologic and prognostic features according to the immunohistochemical expression of Merkel cell polyomavirus large T antigen correlated with viral load","volume":"46","author":"Leroux-Kozal","year":"2015","journal-title":"Hum Pathol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1016\/j.oraloncology.2013.02.003","article-title":"Oral cavity tumors in younger patients show a poor prognosis and do not contain viral RNA","volume":"49","author":"Br\u00e4gelmann","year":"2013","journal-title":"Oral Oncol"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1111\/odi.12178","article-title":"TP53 mutation and human papilloma virus status of oral squamous cell carcinomas in young adult patients","volume":"20","author":"Braakhuis","year":"2014","journal-title":"Oral Dis"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.3233\/CBM-192402","article-title":"Relationship between p16 expression and prognosis in different anatomic subsites of OSCC","volume":"26","author":"Ni","year":"2019","journal-title":"Cancer Biomark"},{"key":"B35","article-title":"Human papillomavirus genotypes and p16 expression in oral leukoplakia and squamous cell carcinoma","volume":"12","author":"Yang","year":"2019","journal-title":"Int J Clin Exp Pathol"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1038\/nature14129","article-title":"Comprehensive genomic characterization of head and neck squamous cell carcinomas","volume":"517","year":"2015","journal-title":"Nature"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1016\/j.oraloncology.2018.06.006","article-title":"Frequent HPV-independent p16\/INK4A overexpression in head and neck cancer","volume":"83","author":"Lechner","year":"2018","journal-title":"Oral Oncol"},{"key":"B38","doi-asserted-by":"publisher","first-page":"691","DOI":"10.1097\/PAS.0000000000001433","article-title":"HPV-positive Squamous Cell Carcinoma of the Larynx, Oral Cavity, and Hypopharynx: Clinicopathologic Characterization With Recognition of a Novel Warty Variant","volume":"44","author":"Rooper","year":"2020","journal-title":"Am J Surg Pathol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.18.00684","article-title":"Human Papillomavirus Testing in Head and Neck Carcinomas: ASCO Clinical Practice Guideline Endorsement of the College of American Pathologists Guideline","volume":"36","author":"Fakhry","year":"2018","journal-title":"J Clin Oncol"},{"key":"B40","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1111\/j.1600-0714.2006.00385.x","article-title":"HPV detection in primary intra-oral squamous cell carcinomas\u2013commensal, aetiological agent or contamination","volume":"35","author":"Boy","year":"2006","journal-title":"J Oral Pathol Med Off Publ Int Assoc Oral Pathol Am Acad Oral Pathol"},{"key":"B41","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1200\/PO.18.00022","article-title":"Predisposing Germline Mutations in Young Patients With Squamous Cell Cancer of the Oral Cavity","author":"Fostira","year":"2018","journal-title":"JCO Precis Oncol"},{"key":"B42","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1038\/nrc2982","article-title":"The molecular biology of head and neck cancer","volume":"11","author":"Leemans","year":"2011","journal-title":"Nat Rev Cancer"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.4149\/neo_2016_013","article-title":"Human papillomavirus and Epstein-Barr virus in nasopharyngeal carcinoma in a non-endemic eastern european population","volume":"63","author":"Svajdler","year":"2016","journal-title":"Neoplasma"},{"key":"B44","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1016\/j.canlet.2016.01.040","article-title":"Global trends in incidence and mortality of nasopharyngeal carcinoma","volume":"374","author":"Tang","year":"2016","journal-title":"Cancer Lett"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1002\/hed.25450","article-title":"Epstein-Barr virus and human papillomaviruses as favorable prognostic factors in nasopharyngeal carcinoma: A nationwide study in Finland","volume":"41","author":"Ruuskanen","year":"2019","journal-title":"Head Neck"},{"key":"B46","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1016\/S0140-6736(19)30956-0","article-title":"Nasopharyngeal carcinoma","volume":"394","author":"Chen","year":"2019","journal-title":"Lancet"},{"key":"B47","doi-asserted-by":"publisher","first-page":"2721367","DOI":"10.1155\/2017\/2721367","article-title":"Detection of Human Papillomavirus Genotypes and Epstein-Barr Virus in Nasopharyngeal Carcinomas at the Korle-Bu Teaching Hospital, Ghana","volume":"2017","author":"Asante","year":"2017","journal-title":"ScientificWorldJournal"},{"key":"B48","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1186\/1750-9378-8-30","article-title":"Oncogenic human papillomavirus-associated nasopharyngeal carcinoma: an observational study of correlation with ethnicity, histological subtype and outcome in a UK population","volume":"8","author":"Robinson","year":"2013","journal-title":"Infect Agent Cancer"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1016\/j.amjoto.2019.06.007","article-title":"HPV status in patients with nasopharyngeal carcinoma in the United States: A SEER database study","volume":"40","author":"Wotman","year":"2019","journal-title":"Am J Otolaryngol"},{"key":"B50","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1002\/ijc.11389","article-title":"A subset of head and neck squamous cell carcinomas exhibits integration of HPV 16\/18 DNA and overexpression of p16INK4A and p53 in the absence of mutations in p53 exons 5-8","volume":"107","author":"Hafkamp","year":"2003","journal-title":"Int J Cancer"},{"key":"B51","doi-asserted-by":"publisher","first-page":"919","DOI":"10.3390\/cancers11070919","article-title":"The Antiviral Agent Cidofovir Induces DNA Damage and Mitotic Catastrophe in HPV-Positive and -Negative Head and Neck Squamous Cell Carcinomas In Vitro","volume":"11","author":"Verhees","year":"2019","journal-title":"Cancers"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1007\/s00262-019-02450-5","article-title":"Potential oncogenic roles of mutant-p53-derived exosomes in the tumor-host interaction of head and neck cancers","volume":"69","author":"Azulay","year":"2020","journal-title":"Cancer Immunol Immunother"},{"key":"B53","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1186\/s40463-018-0302-y","article-title":"The molecular mechanisms of increased radiosensitivity of HPV-positive oropharyngeal squamous cell carcinoma (OPSCC): an extensive review","volume":"47","author":"Liu","year":"2018","journal-title":"J Otolaryngol Head Neck Surg"},{"key":"B54","doi-asserted-by":"publisher","first-page":"15","DOI":"10.21037\/cco.2018.04.05","article-title":"Combinatorial strategies of radiotherapy and immunotherapy in nasopharyngeal carcinoma","volume":"7","author":"Yeo","year":"2018","journal-title":"Chin Clin Oncol"},{"key":"B55","doi-asserted-by":"publisher","first-page":"e00206","DOI":"10.1016\/j.heliyon.2016.e00206","article-title":"Understanding molecular markers in recurrent oral squamous cell carcinoma treated with chemoradiation","volume":"2","author":"Gupta","year":"2016","journal-title":"Heliyon"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1111\/j.1601-0825.2006.01274.x","article-title":"Non-smoking and non-drinking patients with head and neck squamous cell carcinoma: a distinct population","volume":"13","author":"Farshadpour","year":"2007","journal-title":"Oral Dis"}],"container-title":["Frontiers in Oncology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fonc.2020.560434\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,1,20]],"date-time":"2021-01-20T01:57:00Z","timestamp":1611107820000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fonc.2020.560434\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,20]]},"references-count":56,"alternative-id":["10.3389\/fonc.2020.560434"],"URL":"https:\/\/doi.org\/10.3389\/fonc.2020.560434","relation":{},"ISSN":["2234-943X"],"issn-type":[{"value":"2234-943X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,20]]},"article-number":"560434"}}