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PLoS One. 2017 Sep 19;12(9):e0184201. doi: 10.1371/journal.pone.0184201. eCollection 2017.

No evidence for the presence of Epstein-Barr virus in squamous cell carcinoma of the mobile tongue.

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Department of Clinical Sciences/ENT, Umeå University, Umeå, Sweden.
United Arab Emirates University, College of Medicine & Health Sciences, Dept. of Medical Microbiology and Immunology, Tawam Hospital Campus, Al Ain, UAE.
RECAMO, Masaryk Memorial Cancer Institute, Zluty kopec 7, Brno, Czech Republic.
Department of Medical Biosciences, Umeå University, Umeå, Sweden.
University Paris Diderot, INSERM UMRS1162, 27 rue Juliette Dodu, Paris, France.
Department of Neuroscience Reproductive and Dentistry Sciences, University of Naples Federico II, Naples, Italy.
Second University of Naples, Multidisciplinary Department of Medical, Surgical and Dental Specialties, Naples, Italy.
Dipartimento Universitario di Anatomia Patologica, Seconda Universita' Degli Studi di Napoli, Piazza Miraglia, Naples, Italy.


Squamous cell carcinoma of the head and neck (SCCHN) comprises a large group of cancers in the oral cavity and nasopharyngeal area that typically arise in older males in association with alcohol/tobacco usage. Within the oral cavity, the mobile tongue is the most common site for tumour development. The incidence of tongue squamous cell carcinoma (TSCC) is increasing in younger people, which has been suggested to associate with a viral aetiology. Two common human oncogenic viruses, human papilloma virus (HPV) and Epstein-Barr virus (EBV) are known causes of certain types of SCCHN, namely the oropharynx and nasopharynx, respectively. EBV infects most adults worldwide through oral transmission and establishes a latent infection, with sporadic productive viral replication and release of virus in the oral cavity throughout life. In view of the prevalence of EBV in the oral cavity and recent data indicating that it infects tongue epithelial cells and establishes latency, we examined 98 cases of primary squamous cell carcinoma of the mobile tongue and 15 cases of tonsillar squamous cell carcinoma for the presence of EBV-encoded RNAs (EBERs), EBV DNA and an EBV-encoded protein, EBNA-1. A commercially available in situ hybridisation kit targeting EBER transcripts (EBER-ISH) showed a positive signal in the cytoplasm and/or nuclei of tumour cells in 43% of TSCCs. However, application of control probes and RNase A digestion using in-house developed EBER-ISH showed identical EBER staining patterns, indicating non-specific signals. PCR analysis of the BamH1 W repeat sequences did not identify EBV genomes in tumour samples. Immunohistochemistry for EBNA-1 was also negative. These data exclude EBV as a potential player in TSCC in both old and young patients and highlight the importance of appropriate controls for EBER-ISH in investigating EBV in human diseases.

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