Cutaneous human papillomavirus E7 type-specific effects on differentiation and proliferation of organotypic skin cultures.
BACKGROUND
A role for cutaneous human beta-papillomavirus (HPV) types as co-factors in the development of non-melanoma skin cancer has been postulated. Here we have investigated the effects of E7 expression on keratinocyte differentiation, proliferation and cell-cycle proteins in organotypic skin cultures.


METHODS
Recombinant retroviruses containing the E7 genes from cutaneous HPV types 1, 4, 5, 8, 20, 38 and RTRX7 were produced that include types associated with benign and malignant lesions. Adult human primary keratinocytes were transduced with these recombinant retroviruses and differentiated into skin-equivalents using de-epidermalised human dermis.


RESULTS
Expression patterns of the basal keratinocyte marker cytokeratin 14 (CK14) were not altered by any of the viral E7 types analysed. However, expression of the early and late differentiation markers CK10 and involucrin were markedly altered in HPV 1, 4 and 38 cultures. The highest proliferation rates in basal cell layers, as judged by BrdU and Ki67 staining, were observed in HPV 1, 4 and 38 cultures. Interestingly, co-expression of cyclin E and p16(INK4a) within the same cell of the suprabasal cell layers was observed only in cultures generated using E7 of HPV 5 or HPV 8.


CONCLUSIONS
HPV types associated with either benign or malignant lesions perturb keratinocyte proliferation and differentiation in different ways. Moreover, expression of E7 from HPV 5 or HPV 8 seem able to overcome p16(INK4a) induced cell cycle arrest in a subset of keratinocytes.
Westphal
K
K
Department of Dermatology, Venereology and Allergy, Charité, Skin Cancer Center Charité, University Hospital of Berlin, Berlin, Germany.
Akgül
B
B
Storey
A
A
Nindl
I
I
D052685:Alphapapillomavirus / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism / Q000821:virology; D007633:Keratins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D050725:Papillomavirus E7 Proteins / Q000235:genetics / Q000378:metabolism*; D030361:Papillomavirus Infections / Q000378:metabolism / Q000503:physiopathology* / Q000821:virology; D012867:Skin / Q000166:cytology* / Q000378:metabolism / Q000821:virology; D013045:Species Specificity; D055815:Young Adult
D019941:Cyclin-Dependent Kinase Inhibitor p16; D050725:Papillomavirus E7 Proteins; D007633:Keratins