Gulec Cagri, Coban Neslihan, Ozsait-Selcuk Bilge, Sirma-Ekmekci Sema, Yildirim Ozlem, Erginel-Unaltuna Nihan
Istanbul University, Institute for Experimental Medicine, Department of Genetics, Vakif Gureba C. Capa-Fatih, 34104 Istanbul, Turkey.
Exp Cell Res. 2017 Apr 1;353(1):6-15. doi: 10.1016/j.yexcr.2017.02.028. Epub 2017 Feb 24.
ROR-alpha is a nuclear receptor, activity of which can be modulated by natural or synthetic ligands. Due to its possible involvement in, and potential therapeutic target for atherosclerosis, we aimed to identify ROR-alpha target genes in monocytic and endothelial cell lines. We performed chromatin immunoprecipitation (ChIP) followed by tiling array (ChIP-on-chip) for ROR-alpha in monocytic cell line THP1 and endothelial cell line HUVEC. Following bioinformatic analysis of the array data, we tested four candidate genes in terms of dependence of their expression level on ligand-mediated ROR-alpha activity, and two of them in terms of promoter occupancy by ROR-alpha. Bioinformatic analyses of ChIP-on-chip data suggested that ROR-alpha binds to genomic regions near the transcription start site (TSS) of more than 3000 genes in THP1 and HUVEC. Potential ROR-alpha target genes in both cell types seem to be involved mainly in membrane receptor activity, signal transduction and ion transport. While SPP1 and IKBKA were shown to be direct target genes of ROR-alpha in THP1 monocytes, inflammation related gene HMOX1 and heat shock protein gene HSPA8 were shown to be potential target genes of ROR-alpha. Our results suggest that ROR-alpha may regulate signaling receptor activity, and transmembrane transport activity through its potential target genes. ROR-alpha seems also to play role in cellular sensitivity to environmental substances like arsenite and chloroprene. Although, the expression analyses have shown that synthetic ROR-alpha ligands can modulate some of potential ROR-alpha target genes, functional significance of ligand-dependent modulation of gene expression needs to be confirmed with further analyses.
ROR-α是一种核受体,其活性可被天然或合成配体调节。由于其可能参与动脉粥样硬化以及作为潜在的治疗靶点,我们旨在鉴定单核细胞系和内皮细胞系中的ROR-α靶基因。我们在单核细胞系THP1和内皮细胞系HUVEC中对ROR-α进行了染色质免疫沉淀(ChIP),随后进行了芯片平铺分析(ChIP-on-chip)。在对阵列数据进行生物信息学分析之后,我们根据四个候选基因的表达水平对配体介导的ROR-α活性的依赖性进行了检测,并根据ROR-α对启动子的占据情况对其中两个基因进行了检测。ChIP-on-chip数据的生物信息学分析表明,ROR-α与THP1和HUVEC中3000多个基因转录起始位点(TSS)附近的基因组区域结合。两种细胞类型中的潜在ROR-α靶基因似乎主要参与膜受体活性、信号转导和离子转运。虽然在THP1单核细胞中SPP1和IKBKA被证明是ROR-α的直接靶基因,但炎症相关基因HMOX1和热休克蛋白基因HSPA8被证明是ROR-α的潜在靶基因。我们的结果表明,ROR-α可能通过其潜在靶基因调节信号受体活性和跨膜转运活性。ROR-α似乎也在细胞对诸如亚砷酸盐和氯丁二烯等环境物质的敏感性中发挥作用。虽然表达分析表明合成的ROR-α配体可以调节一些潜在的ROR-α靶基因,但基因表达的配体依赖性调节的功能意义需要通过进一步分析来证实。
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