Singh Prashant K, Long Mark D, Battaglia Sebastiano, Hu Qiang, Liu Song, Sucheston-Campbell Lara E, Campbell Moray J
a Departments of Pharmacology & Therapeutics ; Roswell Park Cancer Institute ; Buffalo , NY USA.
Epigenetics. 2015;10(1):40-9. doi: 10.4161/15592294.2014.989088. Epub 2015 Jan 29.
The Vitamin D Receptor (VDR) is a member of the nuclear receptor superfamily and is of therapeutic interest in cancer and other settings. Regulation of microRNA (miRNA) by the VDR appears to be important to mediate its actions, for example, to control cell growth. To identify if and to what extent VDR-regulated miRNA patterns change in prostate cancer progression, we undertook miRNA microarray analyses in 7 cell models representing non-malignant and malignant prostate cells (RWPE-1, RWPE-2, HPr1, HPr1AR, LNCaP, LNCaP-C4-2, and PC-3). To focus on primary VDR regulatory events, we undertook expression analyses after 30 minutes treatment with 1α,25(OH)2D3. Across all models, 111 miRNAs were significantly modulated by 1α,25(OH)2D3 treatment. Of these, only 5 miRNAs were modulated in more than one cell model, and of these, only 3 miRNAs were modulated in the same direction. The patterns of miRNA regulation, and the networks they targeted, significantly distinguished the different cell types. Integration of 1α,25(OH)2D3-regulated miRNAs with published VDR ChIP-seq data showed significant enrichment of VDR peaks in flanking regions of miRNAs. Furthermore, mRNA and miRNA expression analyses in non-malignant RWPE-1 cells revealed patterns of miRNA and mRNA co-regulation; specifically, 13 significant reciprocal patterns were identified and these patterns were also observed in TCGA prostate cancer data. Lastly, motif search analysis revealed differential motif enrichment within VDR peaks flanking mRNA compared to miRNA genes. Together, this study revealed that miRNAs are rapidly regulated in a highly cell-type specific manner, and are significantly co-integrated with mRNA regulation.
维生素D受体（VDR）是核受体超家族的成员，在癌症和其他疾病环境中具有治疗意义。VDR对微小RNA（miRNA）的调控似乎对介导其作用很重要，例如控制细胞生长。为了确定在前列腺癌进展过程中VDR调控的miRNA模式是否以及在多大程度上发生变化，我们在7种代表非恶性和恶性前列腺细胞的细胞模型（RWPE-1、RWPE-2、HPr1、HPr1AR、LNCaP、LNCaP-C4-2和PC-3）中进行了miRNA微阵列分析。为了聚焦于VDR的主要调控事件，我们在用1α,25(OH)2D3处理30分钟后进行了表达分析。在所有模型中，111种miRNA受到1α,25(OH)2D3处理的显著调节。其中，只有5种miRNA在不止一种细胞模型中受到调节，而在这些miRNA中，只有3种在相同方向上受到调节。miRNA调控模式及其靶向的网络显著区分了不同的细胞类型。将1α,25(OH)2D3调控的miRNA与已发表的VDR ChIP-seq数据整合显示，VDR峰在miRNA侧翼区域显著富集。此外，对非恶性RWPE-1细胞的mRNA和miRNA表达分析揭示了miRNA和mRNA的共调控模式；具体而言，鉴定出13种显著的相互调控模式，并且在TCGA前列腺癌数据中也观察到了这些模式。最后，基序搜索分析揭示了与miRNA基因相比，mRNA侧翼VDR峰内的基序富集存在差异。总之，这项研究表明，miRNA以高度细胞类型特异性的方式被快速调控，并且与mRNA调控显著共同整合。
