Genome-wide expression profiling of SGTA knockdown in C4-2B prostate cancer cells. Homo sapiens

Identifying the effect of the co-chaperone SGTA on global androgen receptor transcriptional activity in C4-2B prostate cancer cells with view to further elucidating the broader biological role of SGTA on other signaling pathways within prostate cancer cells Knockdown of SGTA for 72 hours in C4-2B cells significantly altered the expression of approximately 1900 genes in both vehicle and DHT treated cells. The effect of SGTA knockdown was to suppress the expression of approximately 60% of those transcripts. The regulation of 35% of DHT target genes was also affected by SGTA knockdown, with gene-specific effects on basal, or DHT-induced expression, or both. Overall design: C4-2B cells were transfected with 5nM non-specific control siRNA (NS) or with a pool of three commercially avaliable SGTA specific siRNA (SGTA) for 72hrs. Cells were subsequently treated with either ethanol vehicle control or 1nM DHT for 16hr. Total RNA was extracted. Five independent vehicle and 2 DHT siNS and siSGTA samples were hybridized to Affymetrix Human Gene 1.0 ST array chips.

本研究旨在明确共伴侣蛋白（co-chaperone）SGTA对C4-2B前列腺癌细胞中全局雄激素受体（androgen receptor）转录活性的影响，并进一步阐明SGTA在前列腺癌细胞内其他信号通路中的更为广泛的生物学功能。在C4-2B细胞中对SGTA进行72小时基因敲低后，溶剂对照与二氢睾酮（DHT）处理的细胞中均有约1900个基因的表达发生显著改变。SGTA敲低可抑制其中约60%的转录本的表达。此外，SGTA敲低还影响了35%的DHT靶基因的调控，且对这些基因的基础表达、DHT诱导的表达或二者均具有基因特异性的调控作用。实验设计：将C4-2B细胞分别转染5nM非特异性对照小干扰RNA（non-specific control siRNA，简称NS）或三剂市售SGTA特异性小干扰RNA混合池（SGTA），处理72小时；随后分别以乙醇溶剂对照或1nM DHT处理细胞16小时。提取总RNA后，将5份独立的溶剂对照样本、2份siNS转染组DHT处理样本以及2份siSGTA转染组DHT处理样本与Affymetrix人类基因1.0 ST芯片（Affymetrix Human Gene 1.0 ST array）进行杂交。