title: Lack of evidence of human papillomavirus-induced squamous cell carcinomas of the oral cavity in southern Germany.
abstract: OBJECTIVE
  The aim of the present study was to identify HPV-attributable SCC of the oral cavity (OSCC) in a cohort of patients from southern Germany.
  
  
  METHODS
  A sensitive PCR-enzyme immunoassay (EIA) was followed by a more specific in situ hybridization (ISH) to detect high risk human papillomavirus (HPV). An immunohistochemical dual-staining for p16(INK4a) and the proliferation marker Ki-67 was used to assess whether co-expression of p16(INK4a)/Ki-67 is a better surrogate marker for HPV in OSCC than p16(INK4a) alone, based on the hypothesis that combined p16(INK4a) and Ki-67 expression might specifically discriminate oncogene-induced p16(INK4a) expression from cell-cycle arrest-inducing senescence-associated p16(INK4a) expression.
  
  
  RESULTS
  HPV-DNA by PCR-EIA could be detected in 25.1% (69/275) of the tumors, but ISH was negative in all of them. Diffuse p16(INK4a) overexpression was detected in 11 HPV PCR-positive tumors, but also in 6 HPV PCR-negative tumors. p16(INK4a)-expressing cells in diffusely positive tumors co-expressed Ki-67, irrespective of the HPV status. Neither the sole HPV status nor combined HPV/p16(INK4a) status nor the sole p16(INK4a) status was significantly associated with disease free or overall survival, however a trend towards better overall survival of patients whose tumor expressed p16(INK4a) in a focal pattern (=p16(INK4a)-positive/Ki-67-negative cells) compared to no p16(INK4a) expression (p=0.09) was observed.
  
  
  CONCLUSIONS
  Viral DNA can be detected in some tumors by a sensitive PCR, but absence of ISH signals indicates that the HPV-attributable fraction is smaller than estimated from PCR positivity. p16(INK4a)/Ki-67 co-expression is detectable in a fraction of OSCC irrespective of the HPV status.
authors.affiliation: Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; Clinical Cooperation Unit Applied Tumor Biology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 224, 69120 Heidelberg, Germany. Electronic address: miriam.reuschenbach@med.uni-heidelberg.de.
authors.forename: Miriam
authors.initials: M
authors.lastname: Reuschenbach
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany.
authors.forename: Katinka
authors.initials: K
authors.lastname: Kansy
authors.affiliation: Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; Clinical Cooperation Unit Applied Tumor Biology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
authors.forename: Kira
authors.initials: K
authors.lastname: Garbe
authors.affiliation: Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; Clinical Cooperation Unit Applied Tumor Biology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Vinokurova
authors.affiliation: Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; National Center for Tumor Diseases (NCT) Tissue Bank, Im Neuenheimer Feld 221, 69120 Heidelberg, Germany.
authors.forename: Christa
authors.initials: C
authors.lastname: Flechtenmacher
authors.affiliation: Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; National Center for Tumor Diseases (NCT) Tissue Bank, Im Neuenheimer Feld 221, 69120 Heidelberg, Germany.
authors.forename: Csaba
authors.initials: C
authors.lastname: Toth
authors.affiliation: Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; Clinical Cooperation Unit Applied Tumor Biology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
authors.forename: Elena-Sophie
authors.initials: ES
authors.lastname: Prigge
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany.
authors.forename: Oliver C
authors.initials: OC
authors.lastname: Thiele
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Tübingen, Osianderstr. 2, 72076 Tübingen, Germany.
authors.forename: Siegmar
authors.initials: S
authors.lastname: Reinert
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hoffmann
authors.affiliation: Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany; Clinical Cooperation Unit Applied Tumor Biology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
authors.forename: Magnus
authors.initials: M
authors.lastname: von Knebel Doeberitz
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany.
authors.forename: Kolja
authors.initials: K
authors.lastname: Freier
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000032:analysis; D002294:Carcinoma, Squamous Cell / Q000453:epidemiology* / Q000821:virology; D005260:Female; D005838:Genotype; D005858:Germany / Q000453:epidemiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000453:epidemiology* / Q000821:virology; D027383:Papillomaviridae / Q000302:isolation & purification*; D011379:Prognosis
chemical_list: D014408:Biomarkers, Tumor
keywords: DFS; EIA; HPV; HR-HPV; Human papillomavirus; ISH; Ki-67; OS; OSCC; Oral cavity; PCR; SCC; Squamous cell cancer; disease-free survival; enzyme immunoassay; high-risk human papillomavirus; human papillomavirus; in situ hybridization; overall survival; p16(INK4a); polymerase chain reaction; squamous cell cancer; squamous cell cancer of the oral cavity