Immortalization of differentiated human keratinocytes by human papillomavirus (HPV) 16 DNA

Wenjing Zhou, Stephen K. Tyring, Miriam Brysk, Teh Sheng Chan

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Human papillomavirus (HPV) 16 is strongly implicated as having an etiologic role in cervical carcinoma. Human basal keratinocytes have been used in most HPV transformation studies, and transformed cells have exhibited undifferentiated characteristics. However, tumor cells in cervical squamous cell carcinoma are differentiated to certain degrees. Therefore, we hypothesize that HPV may transform not only basal cells but also differentiated keratinocytes. Human keratinocytes isolated from neonatal foreskin were induced to differentiate by treatment with 2 mM Ca2+ for various times (24, 48, 72 and 96 h) and were transfected with HPV 16 DNA. After G418 selection and repeated subculture, HPV 16-transformed cells were derived from each group of the Ca2+-treated and untreated cells. HPV DNA was detected in the transformed cells by polymerase chain reaction (PCR). Southern analysis and two-dimensional gel results indicated that HPV 16 DNA had been integrated into the chromosomes. The HPV 16-transformed cells exhibited an indefinite lifespan and were not tumorigenic in nude mice. They also showed the morphology and expressed the markers of differentiated keratinocytes. This study showed that differentiated human keratinocytes could be immortalized by HPV 16, and this may be a better model for cervical carcinomas in humans.

Original languageEnglish (US)
Pages (from-to)140-152
Number of pages13
JournalJournal of Dermatological Science
Volume13
Issue number2
DOIs
StatePublished - Nov 1 1996

Keywords

  • Carcinogenesis
  • Cervical carcinoma
  • Transformation
  • Tumor virus

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology

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