Human papillomavirus infection and p16INK4a protein expression in vulvar intraepithelial neoplasia and invasive squamous cell carcinoma

Irina Rufforny, Edward J. Wilkinson, Chen Liu, Haizhen Zhu, Michael Buteral, Nicole A. Massoll

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


Objective. Vulvar intraepithelial neoplasia (VIN) is defined histopathologically by distinctive abnormalities of cellular maturation and differentiation. The purpose of this study was to investigate the functional properties of VIN related to expression of p16INK4a protein as well as to detection of human papillomavirus (HPV) type 16 by real-time polymerase chain reaction (RT-PCR) analysis. Methods. A total of 49 vulvar biopsy samples were examined by hematoxylin-eosin staining from benign/reactive lesions, condyloma acuminatum, VIN, and invasive squamous cell carcinoma (SCC). JC8 mouse monoclonal antibodies were used that recognize p16INK4a epitope at a dilution of 1:25. The reaction pattern for p16INK4a was graded in each sample between 0 and 3+. RT-PCR analysis of formalin-fixed paraffin-embedded sections determined positivity for HPV type 16. Results. p16INK4a immunoreactivity was different in VIN 1, VIN 2, VIN 3, and squamous cell carcinoma. Strong expression of p16INK4a protein was observed in 92% (22 of 24) of VIN 2 and VIN 3 lesions and 100% (4 of 4) of invasive SCCs. Two (67%) of 3 VIN 2 lesions, 17 (81%) of 21 VIN 3 lesions, and 4(100%) of 4 SCCs were positive for HPV type 16 by PCR analysis. Two (20%) of 10 VIN 1 lesions were immunoreactive for p16INK4a, with only 1 lesion positive for HPV type 16. No p16INK4a immunoreactivity was observed in any of the benign/reactive and condyloma acuminatum lesions. In addition, none of the benign/reactive or condyloma lesions were positive for HPV type 16 by RT-PCR analysis. Conclusions. Upregulation of INK4a gene occurs in vulvar carcinogenesis. p16INK4a is not a sensitive marker for differentiation of benign vulvar squamous epithelium from condyloma acuminatum or VIN 1 lesions because most VIN 1 lesions are p16INK4a negative. Expression of p16INK4a may aid in the diagnosis of HPV-related lesions and as such may be of value as a surrogate marker in the diagnosis of vulvar premalignant and malignant lesions.

Original languageEnglish (US)
Pages (from-to)108-113
Number of pages6
JournalJournal of Lower Genital Tract Disease
Issue number2
StatePublished - Apr 1 2005
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Obstetrics and Gynecology


  • HPV
  • RT-PCR assay
  • VIN
  • Vulva
  • p16

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