Characterisation of ER-beta functions in prostate cancer (RNA-Seq)

To investigate the effect of a novel ER-beta selective ligand alone and with Enzalutamide on RNA expression in prostate cancer cell lines LNCaP, LNCaP C4-2 and 22Rv1 cells in triplicate were treated with either vehicle, OSU-ERb-12 alone (100 nM) or E2 alone (100nM), Enza (1 microMolar) or the combinations for 12h and RNA-Seq was undertaken. . Sequencing libraries prepared with the TruSeq Stranded Total RNA kit (Illumina Inc), from 1ug total RNA. Alignment of raw sequence reads to the human transcriptome (hg38) was performed via Rsubread and transcript abundance estimates were normalized and differentially expressed genes (DEGs) identified using a standard edgeR pipeline.

本研究旨在探究新型雌激素受体β（ER-beta）选择性配体单独使用，以及联合恩扎卢胺（Enzalutamide，缩写Enza）对前列腺癌细胞系LNCaP、LNCaP C4-2及22Rv1的RNA表达水平的影响。实验设置三次生物学重复，各组细胞分别经溶剂对照、100 nM OSU-ERb-12单独处理、100 nM雌二醇（Estradiol，简称E2）单独处理、1 μM恩扎卢胺单独处理，以及上述药物联合处理12小时后，开展RNA测序（RNA-Sequencing，简称RNA-Seq）。测序文库以1 μg总RNA为起始材料，采用TruSeq链特异性总RNA建库试剂盒（Illumina公司）制备。将原始测序读段比对至人类转录组（hg38）的分析通过Rsubread工具完成，随后对转录本丰度进行归一化处理，并采用标准edgeR分析流程鉴定差异表达基因（DEGs）。