FoxA1 specifies unique androgen and glucocorticoid receptor binding events in prostate cancer cells (mRNA)

We report the androgen receptor recruitment to the chromatin of androgen responsive prostate cancer cell lines, LNCaP-1F5 and VCaP in response to physiological androgen 5a-dihydrotestosterone (DHT) using ChIP-sequencing. We compare the AR recruitment by DHT to that by partial agonist/antagonist cyproterone acetate and mifepristone (RU486) in LNCaP-1F5 cells. We also report the role of glucocorticoid receptor recruitment in presence of dexamethasone (Dex) in androgen responsive prostate cancer cells. The AR and GR cistrome analysis is subsequently compared with gene expression data and RNA Pol II analysis. The ChIP-seq has been performed using AR, GR, RNA Pol II antibodies. Expression profiling by microarray of LNCaP-1F5 cells treated with vehicle, 100 nM 5a-dihydrotestosterone (DHT), 100 nM dexamethasone (Dex), 1000 nM cyproterone acetate (CPA), 100 nM mifepristone (RU486) and combination of 100 nM DHT,100 nM Dex for 24 hours.

本研究采用染色质免疫共沉淀测序（ChIP-sequencing）技术，分析生理浓度雄激素5α-二氢睾酮（5α-dihydrotestosterone, DHT）处理下，雄激素应答型前列腺癌细胞系LNCaP-1F5与VCaP的染色质雄激素受体（androgen receptor, AR）招募情况。本研究同时在LNCaP-1F5细胞中，比较了DHT与部分激动剂/拮抗剂醋酸环丙孕酮（cyproterone acetate, CPA）、米非司酮（mifepristone, RU486）介导的AR招募差异。此外，本研究还探究了地塞米松（dexamethasone, Dex）处理下，雄激素应答型前列腺癌细胞中糖皮质激素受体（glucocorticoid receptor, GR）的招募作用。随后，本研究将AR与GR的顺式调控组（cistrome）分析结果，与基因表达数据及RNA聚合酶II（RNA polymerase II, RNA Pol II）分析结果进行了对比。本研究使用AR、GR及RNA Pol II抗体完成了上述ChIP-seq实验。针对经溶剂对照、100 nM 5α-二氢睾酮、100 nM地塞米松、1000 nM醋酸环丙孕酮、100 nM米非司酮，以及100 nM DHT与100 nM Dex联合处理24小时的LNCaP-1F5细胞，本研究通过微阵列（microarray）完成了表达谱检测。