title: Human papillomavirus in oropharyngeal squamous cell carcinoma: assessing virus presence in normal tissue and activity in cervical metastasis.
abstract: OBJECTIVE
  Human papillomavirus (HPV) has been established as an etiologic and prognostic factor in oropharyngeal squamous cell carcinoma (OPSCC). HPV oncogenesis involves expression of E6/E7 oncoproteins, with downstream p53 degradation and pRb inhibition. Although much research has focused on HPV's oncogenic behavior in primary OPSCC, minimal information exists about HPV in adjacent normal and metastatic tissue.
  
  
  METHODS
  Retrospective cohort study
  
  
  METHODS
  Patient-matched tumor, normal, and metastatic tissue was gathered from 42 OPSCC patients and tested with real-time quantitative polymerase chain reaction (RT-qPCR), in situ hybridization (ISH), and immunohistochemistry (IHC). RT-qPCR was performed using total RNA from fresh-frozen tissues and primers for HPV16 E6, E7, and p16 transcripts. HPV ISH was performed to detect the presence of HPV DNA and IHC to detect p16 protein.
  
  
  RESULTS
  Primary tumor, adjacent normal tissue, and tumor metastasis from 17 OPSCC patients were analyzed. When comparing the presence of HPV16 DNA in tumor, metastatic, and normal tissue by ISH, perfect correlation is found at all subsites (P < .0001). However, active infections determined by HPV16 E6 and E7 expression using quantitative polymerase chain reaction or p16 detection by IHC, were present only in primary and metastatic tissue (P = .0012, E6; P = .02, E7). No such correlation was found in normal tissue when compared to primary or metastatic tissue.
  
  
  CONCLUSIONS
  There is a clear pattern of active HPV expression that correlates to disease course. In HPV-positive patients, all sites including primary, metastatic, and normal tissues are DNA positive. Transcriptionally active infections were detected in primary and metastatic tissues, whereas normal tissues appear to have latent infections.
authors.affiliation: Department of Otorhinolaryngology, Mayo Clinic, Rochester, Minnesota 55905, USA.
authors.forename: Rebecca R
authors.initials: RR
authors.lastname: Laborde
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Janus
authors.forename: Steven M
authors.initials: SM
authors.lastname: Olsen
authors.forename: Vivian W
authors.initials: VW
authors.lastname: Wang
authors.forename: Joaquin J
authors.initials: JJ
authors.lastname: Garcia
authors.forename: Rondell P
authors.initials: RP
authors.lastname: Graham
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Moore
authors.forename: Kerry D
authors.initials: KD
authors.lastname: Olsen
authors.forename: Jan L
authors.initials: JL
authors.lastname: Kasperbauer
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Price
authors.forename: Michael
authors.initials: M
authors.lastname: Berres
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Halling
authors.forename: David I
authors.initials: DI
authors.lastname: Smith
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000032:analysis; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000556:secondary / Q000821:virology*; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D015967:Gene Expression Regulation, Viral; D006258:Head and Neck Neoplasms / Q000175:diagnosis / Q000556:secondary* / Q000821:virology; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis / Q000235:genetics; D009959:Oropharyngeal Neoplasms / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis / Q000235:genetics; D030361:Papillomavirus Infections / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics; D012189:Retrospective Studies; D055815:Young Adult
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012367:RNA, Viral; D012097:Repressor Proteins