Prostate cancer cell line LNCaP treated for 2, 4, 6, 8 h with the synthetic androgen R1881 (1nM)

Time series of the prostate cancer cell line LNCaP, treated for 2, 4, 6 and 8 hours with the synthetic androgen R1881. As control, the cells were cultured for 2, 4, 6 and 8 hours in the presence of the same concentration of solvent (ethanol). With this short treatment time, we aimed to identify mainly direct targets of the androgen receptor. LNCaP has a very low growth rate in steroid stripped medium and resumes growth on addition of androgens. Keywords: Time course The LNCaP cell line was maintained in RPMI media with 10% fetal calf serum and penicillin/streptomycin. Before R1881 treatment, cells were androgen-deprived by culture for 72 h in medium containing 10% dextran-filtered, charcoal-stripped fetal calf serum with a medium replacement after 36 h. After androgen deprivation, the medium was supplemented with R1881 to a final concentration of 1 nM. R1881 was dissolved in ethanol; the final concentration of ethanol in the media was 0.1%. In the control, ethanol was added to the same final concentration. R1881-treated and untreated cells were collected after 2, 4, 6 and 8 h. cDNAs from R1881-treated and control cells were compared directly by hybridization to the same microarray. This was done in duplicate with reversed Cy labeling. In total, eight arrays were performed for four time points (one dye swap per time point).