Fig. 5.

Effect of GWARJD10 on androgen receptor activity. (a) Inhibition of androgen-induced SSAT promoter transcriptional activity by GWARJD10 in LNCaP cells: LNCaP cells were transiently transfected with an SSAT promoter-luciferase reporter vector and treated with 2 nmol/L androgen R1881 or vehicle control with or without 1 or 5μmol/lGWARJD10. Cell lysates were collected after 72hr, the relative light units (RLU) of firefly luciferase activity was measured by a luminometer at 480nm, and the ratio of RLU for androgen treated (+R1881) versus vehicle control (−R1881) cells was determined. Similarly, the ratios of RLU for the LNCaP cells treated with androgen (+R1881) versus vehicle control (−R1881) with 1 and 5μmol/l GWARJD10 were determined. SSAT transcriptional activity was significantly reduced with 5μmol/lof GWARJD10 in the androgen treated cells. N≥3 percondition. ^*P = 0.003 compared to zero dose control. (b) Inhibition of induction of PSA: LNCaP cells in low androgen F1C4 medium were treated with drug GWARJD10 at 5μmol/l (D) and/or androgen R1881 at 2 nmol/l (R). The conditions included: vehicle control (−R, −D), 5μmol/l of GWARJD10 drug (−R, +D), androgen-stimulated (+R, −D), and androgen plus 5μmol/l GWARJD10 (+R, +D). GWARJD10 significantly reduced PSA mRNA under the androgen-deprived condition (P <0.01 for −R, −D compared to −R, +D at all timepoints). As expected, 2nM androgen stimulation led to a significant increase in PSA mRNA (P <0.001 for +R, −D compared to −R, −D at all timepoints). However, GWARJD10 did not affect PSA mRNA levels under the 2nM androgen-stimulated condition, as no difference was observed at any timepoint for +R, −D compared to +R,+D. N= 6 percondition.