In this proposal the investigators will focus on the following objectives: 1) to establish HPV-positive cell lines from human cervix by transfection methodologies as well as by direct culture of biopsies; 2) to investigate the regulation of the response of these cell lines to growth factors, especially TGF-a and TGF-b, using conventional monolayer culture and the collagen raft system; and 3) to colocalize TGF-b and HPV mRNA in tissue biopsies obtained from women at different stages of dysplasia including invasive cancer.The raft system will allow the investigators to test the effects of TGF-a and TGF-b on papillomavirus-containing cells under conditions which more faithfully reflect the human infection. In situ hybridization experiments will reveal whether TGF-b is expressed in abnormal cervical tissues, the patterns of expression and the relationship between TGF-b and HPV-expressing cells.
|Hall, W S; Goto-Mandeville, R; Shih, H A et al. (1997) Molecular analysis of episomal human papillomavirus type 16 DNA in a cervical carcinoma cell line. Virus Res 51:183-95|
|Sears, W L; Goto-Mandeville, R; Mirapuri, M et al. (1995) Effects of 12-O-tetradecanoylphorbol-13-acetate on human papillomavirus type 16-positive keratinocytes at different stages of transformation. Mol Carcinog 13:146-56|
|Braun, L; Mikumo, R; Mark, H F et al. (1993) Analysis of the growth properties and physical state of the human papillomavirus type 16 genome in cell lines derived from primary cervical tumors. Am J Pathol 143:832-44|