Synthetic androgen r1881

The CRPC-derived 22Rv1 (ATCC# CRL-2505), DuCaP, and VCaP (kindly provided by dr. Kenneth J. Pienta, Johns Hopkins, Baltimore, USA), the prostate cancer-derived LNCaP (ATCC# CRL-1740), PC-3 (ATCC# CRL-1435), and the bladder carcinoma-derived 5637 (ATCC# HTB-9) cell lines were maintained as monolayer cultures in Roswell Park Memorial Institute (RPMI)-1640 medium (Invitrogen), supplemented with 2 mM l-glutamine and 10% fetal calf serum (FCS; Sigma). Pancreatic carcinoma MIA-PaCa-2 cells (ATCC# CRL-1420) were grown in Dulbecco’s modified Eagle's medium (DMEM) (Invitrogen) with 4.5 g/ml glucose and 1 mM pyruvate, supplemented with 2 mM l-glutamine and 10% FCS and 2.5% of horse serum (Invitrogen). For hormone stimulation experiments, 0.1 nM of synthetic androgen R1881 (PerkinElmer) was added to the medium in combination with 2 µM enzalutamide (Selleck Chemicals) or 0.2% dimethyl sulfoxide (as a vehicle control). Results were reproduced in at least three independent experiments. All cultures were maintained in a humidified atmosphere at 37 °C and 5% CO₂. Cell lines were authenticated in 2016 using the PowerPlex 21 system (Promega) by Eurofins Genomics (Germany). Cells were frequently tested for Mycoplasma infection, using a Mycoplasma-specific PCR, and propagated for no >6 months or 30 passages after resuscitation from the authenticated stocks.