title: p16 Expression Is Not a Surrogate Marker for High-Risk Human Papillomavirus Infection in Periocular Sebaceous Carcinoma.
abstract: OBJECTIVE
  To evaluate the role of high-risk human papillomavirus (HR-HPV) infection in periocular sebaceous carcinoma (SC) using multiple methods of detection, and to determine whether p16 overexpression is present and can be used as a surrogate marker for HR-HPV.
  
  
  METHODS
  Retrospective observational case series with laboratory investigations.
  
  
  METHODS
  Unstained paraffin sections of 35 cases of periocular SC were analyzed with immunohistochemistry for p16 and subjected to polymerase chain reaction (PCR) for HR-HPV. A subset of 18 lesions that were p16-positive was further studied with a novel method of mRNA in situ hybridization (ISH) for the detection of transcriptionally active HR-HPV, an advanced technique with an enhanced sensitivity and specificity.
  
  
  RESULTS
  The clinical findings were in keeping with those of comparable earlier studies. Strong immunohistochemical p16 positivity (meeting the criterion of >70% nuclear and cytoplasmic staining) was present in 29 of 35 cases of periocular SC (82.9%). The selected 18 p16-positive cases tested were negative for HR-HPV using mRNA ISH. PCR yielded unequivocal results with adequate DNA isolated in 24 cases, 23 of which were negative for HR-HPV. One case was positive for HPV type 16, which was found to be a false positive as collaterally determined by mRNA ISH negativity.
  
  
  CONCLUSIONS
  No evidence was found for HR-HPV as an etiologic agent in the development of periocular SC using multiple modalities to maximize sensitivity and specificity and reduce the limitations of any single test. p16 overexpression is common in periocular SC but unrelated to HR-HPV status. Although p16 may be used as a surrogate marker for HR-HPV status in other tissue sites, this interpretation of p16 positivity is not applicable to periocular SC.
authors.affiliation: David G. Cogan Ophthalmic Pathology Laboratory, Massachusetts Eye & Ear Infirmary, Boston, Massachusetts; Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Stagner
authors.affiliation: Department of Oral and Maxillofacial Pathology, University of the Western Cape and NHLS, Cape Town, South Africa.
authors.forename: Amir H
authors.initials: AH
authors.lastname: Afrogheh
authors.affiliation: David G. Cogan Ophthalmic Pathology Laboratory, Massachusetts Eye & Ear Infirmary, Boston, Massachusetts; Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts. Electronic address: Fred_Jakobiec@meei.harvard.edu.
authors.forename: Frederick A
authors.initials: FA
authors.lastname: Jakobiec
authors.affiliation: Department of Pathology, The New York Eye & Ear Infirmary of Mount Sinai, New York, New York.
authors.forename: Codrin E
authors.initials: CE
authors.lastname: Iacob
authors.affiliation: Department of Ophthalmology, Emory University, Atlanta, Georgia.
authors.forename: Hans E
authors.initials: HE
authors.lastname: Grossniklaus
authors.affiliation: Department of Pathology, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Vikram
authors.initials: V
authors.lastname: Deshpande
authors.affiliation: Molecular Pathology Laboratory, Lancet Laboratories, Johannesburg, South Africa.
authors.forename: Christopher
authors.initials: C
authors.lastname: Maske
authors.affiliation: Department of Medical Biosciences, University of the Western Cape, Cape Town, South Africa.
authors.forename: Donovan C
authors.initials: DC
authors.lastname: Hiss
authors.affiliation: Department of Pathology, Massachusetts General Hospital, and Harvard Medical School, Boston, Massachusetts.
authors.forename: William C
authors.initials: WC
authors.lastname: Faquin
mesh_terms: D018266:Adenocarcinoma, Sebaceous / Q000235:genetics / Q000473:pathology / Q000821:virology*; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D060005:Genotyping Techniques; D061809:Human Papillomavirus DNA Tests; D052162:Human papillomavirus 16 / Q000235:genetics; D052161:Human papillomavirus 18 / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D027383:Papillomaviridae / Q000235:genetics; D030361:Papillomavirus Infections / Q000235:genetics / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012333:RNA, Messenger / Q000235:genetics; D012189:Retrospective Studies; D012626:Sebaceous Gland Neoplasms / Q000235:genetics / Q000473:pathology / Q000821:virology*; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral; D012333:RNA, Messenger