Primary cultures of prostate epithelial cells and their ability to activate carcinogens.

Martin, Francis L orcid iconORCID: 0000-0001-8562-4944, Cole, K. J., Muir, Gordon H., Kooiman, G. G., Williams, J. A., Sherwood, R. A., Grover, Philip L. and Phillips, D. H. (2002) Primary cultures of prostate epithelial cells and their ability to activate carcinogens. Prostate Cancer and Prostatic Diseases, 5 (2). pp. 96-104. ISSN 1365-7852

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Differences in the incidence of prostate cancer (CaP) amongst different migrant populations point to causative agents of dietary and/or environmental origin. Prostate tissues were obtained following transurethral resection of the prostate (TURP) or radical retropubic prostatectomy. After surgery, TURP-derived or tumour-adjacent tissue fragments were minced in warm PFMR-4A medium (37?C) and suspensions pipetted into collagen-coated petri dishes. Non-adherent material was removed by washing with fresh medium after 12 h. Adhered cells subsequently reacted positively with monoclonal antibodies to prostate specific antigen (PSA). PSA was also detected in the medium. The genotoxicities of the chemical carcinogens 2-amino-1-methyl-6-phenylimidazo4,5-b pyridine (PhIP), its N-hydroxy metabolite (N-OH-PhIP) and benzoapyrene (BaP) in adherent cell populations from different donors (n=8) were examined. Cells were treated in suspension for 30 min at 37?C in the presence of the DNA repair inhibitors hydroxyurea (HU) and cytosine arabinoside (ara-C). DNA single-strand breaks were detected in cells by the alkaline single cell-gel electrophoresis ('Comet') assay and quantified by measuring comet tail length (CTL) in m. All three carcinogens induced dose-related increases in CTLs (P{\ensuremath{<}}0.0001) in cells from four donors 24 h post-seeding. However, in cells from a further two donors the genotoxic effects of PhIP, N-OH-PhIP and BaP were much less apparent after 48 h than after 24 h in culture. After 96 h in culture, cells from these donors appeared to be resistant to the comet-forming activity of the compounds. However, BaP-DNA adducts were still measurable by 32P-postlabelling for up to 14 days following a 24-h exposure to 50 M BaP in adhered cells from another two donors. This study shows that primary cultures of cells derived from the prostate can activate members of two classes of chemical carcinogens. Further development may provide a robust model system in which to investigate the aetiology of CaP.

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