Transformation of human neonatal prostate epithelial cells by strontium phosphate transfection with a plasmid containing SV40 early region genes.
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 beta 1 (TGF-beta 1) and epidermal growth factor in clonal growth assays. In contrast, a prostatic carcinoma cell line (PC-3) is inhibited by TGF-beta 1. This serum-free system and immortalized transfected clones will be useful for studying the action of putative prostatic carcinogens and tumor-promoting agents.
Kaighn
M E
ME
Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892.
Reddel
R R
RR
Lechner
J F
JF
Peehl
D M
DM
Camalier
R F
RF
Brash
D E
DE
Saffiotti
U
U
Harris
C C
CC
D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival; D002472:Cell Transformation, Viral; D002869:Chromosome Aberrations; D006801:Humans; D007231:Infant, Newborn; D007621:Karyotyping; D008297:Male; D010710:Phosphates / Q000494:pharmacology*; D011467:Prostate / Q000473:pathology / Q000648:ultrastructure*; D011498:Protein Precursors; D011506:Proteins / Q000494:pharmacology; D013539:Simian virus 40; D013324:Strontium / Q000494:pharmacology*; D014162:Transfection; D016212:Transforming Growth Factor beta
D010710:Phosphates; D011498:Protein Precursors; D011506:Proteins; D016212:Transforming Growth Factor beta; C052774:strontium phosphate; D013324:Strontium