Figure 6.

Microarray profiling of prostate stem/progenitor cells following short-term E2 treatment. (A) Microarray heat map of day 7 prostaspheres (PSs) treated with vehicle or 10 nM E2 for 3 h revealed a set of genes modulated by short-term E2 exposure compared with controls. Results are shown from three biologic repeats. (B) Illumina microarray genes showing more than twofold upregulation in day 7 PSs following treatment with 10 nM E2 for 3, 6, or 18 h relative to control. Numbers represent average of n = 3. (C) PSs were exposed to 10 nM E2 dendrimer (impermeable to nuclear membrane) for 24 h and qRT-PCR validated increased expression of several genes that were upregulated more than twofold in the gene expression microarray experiment (n = 3) *P < 0.05; ^#P < 0.005. (D) A subset of the microarray genes upregulated more than twofold by 3 h E2 exposure were validated by qRT-PCR in day 7 PSs treated with 10 nM E2 for 3 h with BGN, MAF, FOSB, and FOXQ1 significantly greater than vehicle controls. Increased BGN, MAF, and FOXQ1 expression was attenuated by ICI pretreatment. Elevated BGN, FOSB, and FOXQ1 expression was also blocked by LY294002 (LY) or U0126 (UO) pretreatment, whereas increased MAF expression was only blocked by LY294002 (n = 7). *P < 0.005 vs vehicle; ^#P < 0.05 vs E2. (E) qRT-PCR showed that cyclin gene expression (CCND1, CCNA1) was significantly increased by 3 h E2 exposure compared with vehicle, and the effect was blocked upon pretreatment with ICI, LY294002, or U0126 (n = 7). *P < 0.05; ^#P < 0.005.