Genome-wide expression profiling of Hic-5 knockdown in PshTert-AR human myofibroblasts

Identifying the effect of the co-regulator Hic-5 (TGFB1I1) on global androgen receptor transcriptional activity in PShTert-AR prostate fibroblast cells with view to further elucidating the broader biological role of Hic-5 on fibroblast spceific androgen signaling. Knockdown of Hic5 for 48 hours in PShTert-AR cells significantly altered the androgen regulation of approximately 1347 genes. The effect of Hic-5 knockdown was to suppress androgen regulation of approximately 85% of those transcripts, by either reducing androgen mediated repression or activition of gene expression. PShTert-AR cells were transfected with 10nM non-specific control siRNA (NS) or with commercially avaliable Hic-5 specific siRNA (siHic5) for 24hrs. Cells were kept in 5% hormone stripped FBS RPMI for a further 24 hours and subsequently treated with either ethanol vehicle control or 10nM DHT for 16hr. Total RNA was extracted. Nine independent vehicle and 9 DHT siNS and siHic5 samples were pooled into three groups of vechicle and DHT treated siNS and siHic5 samples and hybrydised to Affymetrix Human Gene 1.0 ST array chips.