Figure 4

PCA3 expression is upregulated by androgen and modulates the transcription of androgen-regulated genes. (A) PCA3 expression was evaluated in LNCaP cells by qRT-PCR after treatment with 100 nM of DHT or 100 nM DHT plus 100 nM flutamide during a 48-h time course, as described in the Methods section. PCA3 relative expression was determined compared to LNCaP cells treated with ethanol, which was the control vehicle. Error bars +/- SD. (B) Relative RNA quantification of PCA3, AR and androgen-regulated genes (TMPRSS2, NDRG1, GREB1, PSA, AR, FGF8, CdK1, CdK2, and PMEPA1) in LNCaP cells treated with 100 nM of dihydrotestosterone (DHT) for 36 h, compared to cells treated with the control vehicle (ethanol), as described in the Methods section. Bar graphs show the average transcript levels of each gene tested, by qRT–PCR analysis of three independent RNA samples prepared following the treatment of LNCaP cells with DHT or ethanol only. Error bars +/- SD. (C) Relative RNA levels of PCA3, AR, and androgen-regulated genes 36 h after LNCaP cells were transfected with siPCA3, compared to LNCaP/siSCr transfected cells. Error bars +/- SD. (D) Relative RNA levels of PCA3, AR, and androgen-regulated genes 36 h after LNCaP cells were transfected with siPCA3 simultaneously with treatment with 100 nM DHT, compared to LNCaP cells transfected with siScr simultaneously with treatment with 100 nM DHT, as described in the Methods section. 18S RNA was used as a constitutive gene in all these assays. Data are represented as mean ± SD. All experiments were biological replicates repeated a minimum of three times. *, p < 0.01, in comparison to scrambled-siRNA (siScr) treated cells.