Targeted disruption of the epidermal growth factor receptor inhibits development of papillomas and carcinomas from human papillomavirus-immortalized keratinocytes

C. D. Woodworth, D. Gaiotti, E. Michael, Laura A. Hansen, M. Nees

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The epidermal growth factor receptor (EGF-R) is frequently overexpressed in human papillomavirus (HPV)-associated dysplasias and carcinomas, implying that it is important for the progression of keratinocytes to malignancy. We used mice with a targeted disruption of the EGF-R gene to directly examine its role in cell immortalization and tumor development. Epidermal keratinocytes were cultured from EGF-R knockout, heterozygous, and wild-type mice, infected with retroviruses encoding HPV-16 E6 and E7 oncogenes, and grafted to nude mice. E6/E7 induced immortalization of EGF-R wild-type cells 5-fold more efficiently than null cells. Immortal EGF-R null cells grew more slowly, achieved a lower saturation density, and were more sensitive to apoptosis than the immortalized wild-type or heterozygous cells. Analyses using cDNA expression arrays showed that EGF-R null cells expressed increased levels of RNAs encoding p21waf and insulin-like growth factor-binding protein-2. EGF-R-positive immortal keratinocytes formed papillomas in 17% (15 of 90) of skin grafts, and seven grafts progressed to squamous carcinoma after 6-12 months. EGF-R null keratinocytes did not form papillomas, but 1 of 96 grafts progressed to a squamous carcinoma after 1 year. However, treatment with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate induced tumors in 18 and 35% of grafts containing EGF-R null or EGF-R-positive cells, respectively. Transduction with an activated v-Haras gene, which signals downstream of the EGF-R, induced rapidly growing carcinomas in all grafts regardless of EGF-R genotype. These results directly show that the EGF-R is important, but not essential, for immortalization by HPV and for progression of immortal cells to papillomas and carcinomas.

Original languageEnglish
Pages (from-to)4397-4402
Number of pages6
JournalCancer Research
Volume60
Issue number16
StatePublished - Aug 15 2000
Externally publishedYes

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Papilloma
Keratinocytes
Epidermal Growth Factor Receptor
Carcinoma
Null Lymphocytes
Transplants
Squamous Cell Carcinoma
Insulin-Like Growth Factor Binding Protein 2
erbB-1 Genes
Neoplasms
Human papillomavirus 16
Tetradecanoylphorbol Acetate
Retroviridae
Oligonucleotide Array Sequence Analysis
Oncogenes
Nude Mice
Carcinogens
Genotype
RNA
Apoptosis

All Science Journal Classification (ASJC) codes

  • Cancer Research
  • Oncology

Cite this

Targeted disruption of the epidermal growth factor receptor inhibits development of papillomas and carcinomas from human papillomavirus-immortalized keratinocytes. / Woodworth, C. D.; Gaiotti, D.; Michael, E.; Hansen, Laura A.; Nees, M.

In: Cancer Research, Vol. 60, No. 16, 15.08.2000, p. 4397-4402.

Research output: Contribution to journalArticle

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