Epidermal cancer associated with expression of human papillomavirus type 16 E6 and E7 oncogenes in the skin of transgenic mice.

Abstract

Certain "high-risk" anogenital human papilomaviruses (HPVs) have been associated with the majority of human cervical carcinomas. In these cancers, two papillomaviral genes, E6 and E7, are commonly expressed. In this study we provide evidence that expression of the E6 and E7 genes from the high-risk HPV-16 in the skin of transgenic mice potentiated the development of preneoplastic lesions, and a high percentage of these epidermal lesions subsequently devel- In Situ Hybridization. In situ hybridizations were performed as described (12). The cRNA probes for type I epidermal-specific transglutaminase and al type IV collagen were synthesized in vitro from the template DNAs, pTG13 (13) and pCIV 1225 (14), respectively, using UTP [a-35S]. The E6 and E7 open reading frames from HPV-16 were independently cloned into pGEM3Z and each was used to generate cRNA probe, incorporating UTP [a-35S] and CTP [a-35S], and then combined for hybridization. Slides were exposed for 7 days at 4°C in the dark before developing.Single-Strand Conformation Polymorphism (SSCP) Analysis. For SSCP analysis, RNA/PCR products were generated using [a-32PJdATP during amplification, and products were resolved by gel electrophoresis using 0.5 x MDE gel mix with 0.5x Tris/borate/EDTA buffer and 10% glycerol (AT Biochem, Malvern, PA). RESULTS Incidence of Abnormal Skin and Skin Tumors in aAHPV16E6/E7 Transgenic Mice. In characterizing the patterns of transgene expression in tissues from neonatal aAHPV16E6/E7 transgenic mice, we noted expression of E6 and E7 in several nonlenticular tissues, particularly in one line of mice (11). Upon breeding this line of mice, line 19, we noted the frequent appearance of abnormal-looking skin, appearing at ==3 months of age. The incidence of abnormal skin was higher and appearance was earlier in mice homozygous for the transgene than in hemizygous mice (Table 1, 48% Abbreviations: HPV, human papillomavirus; SSCP, single-strand conformation polymorphism; PCNA, proliferating cell nuclear antigen; FITC, fluorescein isothiocyanate.§To whom reprint requests should be addressed.5583

Strong synergy between mutant ras and HPV16 E6/E7 in the development of primary tumors

“…Expression of the v-Ha-ras oncogene in epidermal cells has been shown to render mice susceptible to papilloma development following a 'second event' such as chemical promotion or a mild wounding (Bailleul et al, 1990;Leder et al, 1990). Similarly, expression of the HPV16 E6/E7 oncogenes in epithelial cells can render mice susceptible to the development of squamous cell neoplasms, but the penetrance of neoplasms varies greatly depending upon other genetic predispositions of the mouse strain used (Drinkwater and Bennett, 1991;Lambert et al, 1993;Arbeit et al, 1994;Coussens et al, 1996;Melero et al, 1997). The transgenic C57BL/6 lineage we used does not develop tumors even though E6/E7 is expressed at detectable levels (Melero et al, 1997).…”

Profiling of transcripts and proteins modulated by the E7 oncogene in the lung tissue of E7-Tg mice by the omics approach

“…The lungs of 3-and 7-month-old E7 male mice were removed and compared with those of non-Tg mice, revealing that there was no significant difference between the two groups (data not shown). It has previously been demonstrated that ectopic expression of HPV-16 E6 and E7 in the skin of one HPV-16 transgenic mouse lineage can induce squamous cell carcinomas in 25% of the adult mice (16). However, Greenhalgh et al also directed expression of HPV-16 E6 and E7 together to the epidermis using a suprabasal layer-specific K1 promoter.…”

Interaction of the fork head domain transcription factor MPP2 with the human papilloma virus 16 E7 protein: enhancement of transformation and transactivation

“…Third, each of the transforming proteins can cooperate with an activated form of the oncoprotein Ras in the transformation of primary rat embryo cells (Chesters and McCance, 1989;Crook et al, 1988Crook et al, , 1989DiPaolo et al, 1989;Matlashewski et al, 1987;Phelps et al, 1988). Finally, transgenic mice expressing E7 in epithelial and thyroid cells develop hyperplasias and carcinomas (Auewarakul et al, 1994;Greenhalgh et al, 1994;Herber et al, 1996;Lambert et al, 1993;Ledent et al, 1995).…”