The 3-N-alkyloxyestradiol derivative, 3-[2-diisopro-pylamino]-ethoxy-D1,3,5 (10)-estrien-17-one (DI) was synthesized and provided by Dr. John Cooperwood (Florida A&M University, Tallahassee, FL, USA). Propidium iodide, DMSO (dimethyl sulfoxide), D-Glucose and Ethanol was obtained from Sigma-Aldrich (St. Louis, MO, USA). RPMI 1640, penicillin-streptomycin antibiotic solution (10,000 UI/mL), 0.25% Trypsin-EDTA solution and phosphate buffer (PBS) were all obtained from Invitrogen/Life Technologies (Carlsbad, California, USA). fetal bovine serum (FBS) was obtained from Atlanta Biologicals (Flowery Branch, GA, USA). Quick Start™ Bradford Reagent, 1X (BioRad, Hercules, CA) and Ready 2-D Starter Kit were obtained from BioRad (Hercules, CA, USA).
PC-3 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and penicillin-streptomycin antibiotic solution (10,000 UI/mL), at 37°C in a 5% CO2 humidified atmosphere. A stock solution was prepared by dissolving 3-[2-diisopropylamino]-ethoxy-D1,3,5(10)-estrien-17-one (DI) in 100% DMSO (Sigma-Aldrich) with subsequent dilutions made in cell culture medium. In our experimental conditions, PC-3 cells were treated for 48 h with DI. Cells were detached with 0.25% Trypsin-EDTA solution washed and collected.
PC-3 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and penicillin-streptomycin antibiotic solution (10,000 UI/mL), at 37°C in a 5% CO2 humidified atmosphere. A stock solution was prepared by dissolving 3-[2-diisopropylamino]-ethoxy-D1,3,5(10)-estrien-17-one (DI) in 100% DMSO (Sigma-Aldrich) with subsequent dilutions made in cell culture medium. In our experimental conditions, PC-3 cells were treated for 48 h with DI. Cells were detached with 0.25% Trypsin-EDTA solution washed and collected.