Identification of the GATA2 transcriptome in the AR-V positive castrate-resistant prostate cancer cell line CWR22Rv1

To interrogate the extent of overlap between GATA2 and AR-V regulated transcriptomes in castrate-resistant prostate cancer cells, we assessed global gene expression changes in GATA2-depleted CWR22Rv1 cells by microarray analysis using the Illumina Human HT-12 BeadChip array and compared GATA2 regulated genes to a previously generated AR-V regulated genset (Jones et al., 2015). We identified that AR-V transcriptional activity is dependant upon GATA2 at a subset of its target genes. GATA2 also regulated the expression of c-Myc and E2F1 regulated transcriptional targets amongst other oncogenic transcription factors. Transcripts regulated by GATA2 were highly enriched in the cell cycle. CWR22Rv1 cells were transfected with either a non-silencing scrambled control siRNA (siSCR) or a GATA2 targeting siRNA (siGATA2) 48 hours prior to harvest using Lipfectamine RNAiMAX (Invitrogen) to a final concentration of 25 nM in steroid depleted conditions. Cells were grown for 72 hours in steroid depleted conditions total -/+ 10 µM enzalutamide for 24 hours before RNA extraction using TRIzol (Life Technologies) and hybridization of triplicate samples to an Illumine HT-12 v4 BeadChip array.