Chronic IL-1 Exposed AR+ PCa Cell Lines Show Conserved Loss of IL-1 Sensitivity and Evolve Both Conserved and Unique Differential Gene Expression Profiles

AR+ PCa cell lines show conserved activation of canonical IL-1 intracellular signaling cascades, including repression of androgen receptor mRNA levels, in response to acute IL-1 treatment and show conserved loss of sensitivity to prolonged (chronic) IL-1 intracelluar signaling, including the restoration of the androgen receptor. Thus, chronic IL-1 exposure selects for cells that have a growth advantage against cytotoxic/static inflammation, including restoration of constitutive AR expression. Overall design: MDA-PCa-2b cells were chronically cultured in 0.5 ng/ml IL-1 alpha or IL-1 beta (in HPC1 + 20% FBS) for 4 months. IL-1 is cytotoxic/static, so the viable, proliferating colonies that emerged after 4 months in IL-1 alpha or IL-1 beta were expanded to generate the IL-1a subline (MDA-as1) and the IL-1b subline (MDA-ßs1). MDA-PCa-2b parental cells were cultured in vehicle (0.1% BSA in 1xPBS) in HPC1 + 20% FBS. For basal expression and acute IL-1 treatment expression studies, MDA-PCa-2b parental, MDA-as1 and MDA-ßs1 cells were cultured in vehicle (0.1% BSA in 1xPBS) in HPC1 + 20% FBS for 3 days and the MDA-PCa-2b parental cells were treated with 25 ng/ml IL-1 alpha or IL-1 beta in HPC1 + 20% FBS for 3 days. Total RNA was isolated from 3 biological replicates for RNA sequencing.