Lncap cells

Human, castration-resistant prostate tumor cell lines PC3 and DU145 and castration-sensitive LNCaP cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ). The resistant sublines were derived from the Resistant Cancer Cell Line (RCCL) collection [12 (link)]. Cells were grown and subcultured in RPMI 1640 medium (Gibco/Invitrogen, Karlsruhe, Germany) and supplemented with 10% fetal calf serum (FCS) (Gibco/Invitrogen, Karlsruhe, Germany), 2% HEPES buffer (Sigma-Aldrich, Darmstadt, Germany), 2% glutamine (Gibco/Invitrogen, Karlsruhe, Germany), and 1% penicillin/streptomycin (Gibco/Invitrogen, Karlsruhe, Germany) at 37 °C in a humidified, 5% CO₂ incubator.

LNCaP cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). Cells were grown in Dulbecco's Modified Eagle Medium (DMEM; Pan Biotech, Aidenbach, Germany) supplemented with 10% fetal bovine serum, 1% glutamine and 1% penicillin/streptomycin (all PAN-Biotech) and plasmocin (InvivoGen, San Diego, USA). PC3 prostate cancer cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (ThermoFisher, Waltham, USA), supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (both ThermoFisher). Identity of PC3 cells in-house was verified by cell line authentication (Multiplexion, Heidelberg, Germany). Cells were incubated at 37 °C and 5% CO₂ and seeded 96 h before further analysis of anti-androgen effects. After 24 h, a medium change was carried out and anti-hormones added to half of the cells for 72 h of treatment; 48 h after seeding, anti-hormones were added to the other half for the 48 h determination. Different concentrations of abiraterone acetate (1, 2.5, 5 and 10 μM) and VPC-13566 (1, 10, 50, 100, 200 nM and 1, 2.5, 5, 10, 15 μM) dissolved in DMSO were used (all Merck, Darmstadt, Germany). Cells incubated for 72 h with DMSO were used for control.