Lncap

PCa cells lines LNCaP (CRL-1740), and second-generation LNCaP subline: C4-2 (CRL-3314) and C4-2B (CRL3315) (B for Bone Metastatic) [16 (link)] were purchased from American Type Culture Collection (ATCC). LNCaP cells were cultured in Roswell Park Memorial Institute medium containing D-Glucose (4.5 g/L), HEPES Buffer (2383 g/L), L-Glutamine, Sodium Bicarbonate (1.5 g/L), sodium pyruvate (110 mg/L) (RPMI, A1049101, Lonza, Levallois-Perret, France) and supplemented with 10% fetal bovine serum (FBS) (CVF5VF00-01, Eurobio, Les Ulis, France) and 1% penicillin-streptomycin (PS). C4-2 and C4-2B cells are cultured in RPMI medium (BE12-702F, Lonza, Levallois-Perret, France) supplemented with 5% FBS and 1% PS. Cells were maintained in a 37 °C humidified incubator with 5% CO².
The non-steroidal AR inhibitor, Enza (PHB00235) and the SK3 positive modulator CyPPA (Cyclohexyl-[2-(3,5-dimethyl-pyrazol-1-yl)-6-methyl-pyrimidin-4-yl]-amine) were purchased from Sigma-Aldrich (St-Quentin Fallavier, France). 1-Ohexadecyl-2-O-methyl-sn-glycero-3-lactose (Ohmline) was synthetised as previously described [17 (link)]. For one-week treatments, cells were treated 3 times per week with Enza, Enza+Ohmline or CyPPA.

The prostate carcinoma cell lines PC3 and LNCaP and breast carcinoma cell lines MCF7 and SKBR3 were obtained from ATCC (Manassa, VA, USA). The PC3-9 cell line^{24 (link)}, a sub-clone of the PC3 cell line, was kindly provided by Immunicon (Huntingdon Valley, PA, USA). PC3, PC3-9 and LNCaP were cultured in RPMI1640 (Lonza), MCF7 and SKBR3 in DMEM (Lonza), all supplemented with 10% FBS (Sigma) and 1% Pen/Strep (Lonza). Cells were cultured at 37° in a humid atmosphere and trypsinized when reaching 70–80% confluence using 0.05% trypsin–EDTA (Gibco, Life Technologies, Waltham, MA, USA).