Targeting Jak2-Stat5a/b signaling suppresses growth of enzalutamide-resistant prostate cancer in vitro and in vivo. Homo sapiens

We used cDNA microarray analysis to identify the Stat5a/b transcriptome in AR-depleted CWR22Pc cells. We sequentially depleted AR protein (AR knockdown) followed by Stat5a/b (Stat5a/b knockdown) using lentiviral shRNA in CWR22Pc cells. CWR22Pc cells were transduced with scramble shRNA (shCtrl), shAR or shStat5a/b at the start of the experiment (day 0), followed by transduction with a second lentiviral shRNA, either shCtrl or shStat5a/b, 3 days later (day 3). CWR22Pc cells were harvested on day 6 for analysis of gene expression profiles. Overall design: CWR22Pc cells were sequentially transduced three days apart (on days 0 and 3) with 200-250 µl of lentiviral scramble control shRNA (shCtrl), shRNA-AR (shAR) or shRNA-Stat5a/b (shStat5a/b) in the presence of polybrene (1:1000) as follows: 1) shCtrl>shCtrl (control); 2) shAR>shCtrl (AR knockdown alone); 3) shStat5a/b>shCtrl (Stat5a/b knockdown alone); 4) shAR>shStat5a/b (sequential AR knockdown followed by Stat5a/b knockdown). On day 6, cells were harvested, total RNA was isolated from cells in each treatment group as non-pooled triplicates and samples were prepared for cDNA microarray analysis with Affymetrix GeneChip Human Gene 2.0 ST arrays.