This report describes the development and characterization of an epithelial cell line (BPH-1) from human prostate tissue obtained by transurethral resection. Primary epithelial cell cultures were immortalized with SV40 large T antigen. One of the isolated clones was designated BPH-1. These cells have a cobblestone appearance in monolayer culture and are non-tumorigenic in nude mice following subcutaneous injection or subrenal capsule grafting. They express the SV40 large T antigen and exhibit increased levels of p53, as determined by immunocytochemistry. Cytogenetic analysis by G-banding demonstrated an aneuploid karyotype with a modal chromosome number of 76 (range 71 to 79, n = 28) and 6 to 8 marker chromosomes. Some structurally rearranged chromosomes were observed, but the Y chromosome was normal. The expressed cytokeratin profile was consistent with a prostatic luminal epithelial cell. This profile was the same as that of primary prostatic epithelial cultures from which the BPH-1 cells were derived. In serum-free culture in plastic dishes epidermal growth factor (EGF), transforming growth factor (TGF)-alpha, fibroblast growth factor (FGF) 1 (aFGF), and FGF 7 (KGF) induced increased proliferation in these cells whereas FGF 2 (bFGF), TGF-beta 1, and TGF-beta 2 inhibited proliferative activity. Testosterone had no direct effect on the proliferative rate of BPH-1 cells. 5 alpha-Reductase, 3 alpha-hydroxysteroid oxidoreductase, and 17 beta-hydroxy-steroid oxidoreductase activities were detected in BPH-1 cells. Expression of androgen receptors and the secretory markers, prostate specific antigen and prostatic acid phosphatase, were not detectable by immunocytochemistry, biochemical assay, or RT-PCR analysis.
SUMMARY
Factors controlling the synthesis of androstenedione in the human adrenal gland were investigated in a kinetic study of the enzymes catalysing the conversion of 17α-hydroxyprogesterone and dehydroepiandrosterone to the hormone. Both reactions are associated with the microsomal fraction of the adrenal cell. 17α-Hydroxyprogesterone is converted to androstenedione by a NADPH-dependent 17-desmolase whereas there is an obligatory requirement for NAD+ for the conversion of dehydroepiandrosterone to the hormone. None of the other cofactors investigated inhibited or enhanced the enzymic conversions.
The side-chain cleavage of 17α-hydroxyprogesterone was competitively inhibited by the precursors of the substrate, pregnenolone, progesterone and 17α-hydroxypregnenolone. Other C19 or C18 compounds known to be synthesized by the human adrenal gland were found to be ineffective in modifying the enzyme action. The conversion of dehydroepiandrosterone to androstenedione was non-competitively inhibited by oestrone and oestradiol-17β. No C21 compounds were found to affect the reaction. 5α-Dihydrotestosterone (17β-hydroxy-5α-androstan-3-one) inhibited both enzymic conversions competitively but the inhibition constants observed were so high that it is unlikely that this finding has any relevance to the control of androstenedione synthesis in vitro.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.