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Transformation of Human Neonatal Prostate Epithelial Cells by Strontium Phosphate Transfection with a Plasmid Containing SV40 Early Region Genes

Laboratory of Human Carcinogenesis [M. E. K., R. R. R., J. F. L., D. E. B., C. C. H.] and Laboratory of Experimental Pathology [U. S.], Division of Cancer Etiology, National Cancer Institute, Bethesda, Maryland 20892; Developmental Therapeutics Program [R. F. C.], Division of Cancer Treatment, National Cancer Institute, Frederick, Maryland 20701; and Division of Urology [D. M. P.], Stanford University Medical Center, Stanford, California 94305-5118

Neonatal human prostatic epithelial cells (NP-2s) were transfected by strontium phosphate coprecipitation with a plasmid (pRSV-T) containing the SV40 early region genes. The cells transfected with pRSV-T, but not the sham-transfected controls, formed rapidly growing, multilayered colonies within 2 weeks at a frequency of 1 x 10^-4 in a serum-free medium (P4-8F). In all, 28 colonies of transformed cells were isolated. Three of these have been cultured for a sufficient length of time to show that their growth potentials are well beyond that of the normal progenitor cells (NP-2s). There is also little or no indication of the culture "crisis" commonly seen in SV40-transformed cells in these transfected lines. All contain cytokeratins and SV40 T-antigen as revealed by immunofluorescence, have ultrastructural features of epithelial cells, and are pseudodiploid. None have produced tumors within 1 year after s.c. injection into nude mice. The transformed as well as the parental NP-2s cells require bovine pituitary extract for growth in serum-free medium and are stimulated by transforming growth factor ß₁ (TGF-ß₁) and epidermal growth factor in clonal growth assays. In contrast, a prostatic carcinoma cell line (PC-3) is inhibited by TGF-ß₁. This serum-free system and immortalized transfected clones will be useful for studying the action of putative prostatic carcinogens and tumor-promoting agents.

¹ To whom correspondence and requests for reprints should be addressed.

² Present address: Children's Medical Research Foundation, P. O. Box 61, Camperdown, N. S. W. 2050, Australia.

Received 9/21/88. Revised 2/17/89. Accepted 2/27/89.

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