Epigenetics and Cancer Research Laboratory, Biochemistry and Molecular Biology Group, Department of Life Science, National Institute of Technology, Rourkela, Odisha 769008, India.
Epigenetics and Cancer Research Laboratory, Biochemistry and Molecular Biology Group, Department of Life Science, National Institute of Technology, Rourkela, Odisha 769008, India.
Gene. 2019 Jul 15;705:22-35. doi: 10.1016/j.gene.2019.04.046. Epub 2019 Apr 18.
Mixed-lineage leukaemia 1 (MLL1) enzyme plays major role in regulating genes associated with vertebrate development. Cell physiology and homeostasis is regulated by microRNAs in diverse microenvironment. In this investigation we have identified conserved miR-193a target sites within the 3'-UTR of MLL1 gene transcript. Utilizing wild type and mutated 3'-UTR constructs and luciferase reporter assays we have clearly demonstrated that miR-193a directly targets the 3'-UTR region of the MLL1 mRNA. Ectopic expression of miR-193a modulated global H3K4 mono-, di- and tri-methylation levels and affects the expression of CAV1, a gene which is specifically modulated by H3K4me3. To determine the implications of this in vitro finding in aberrant physiological conditions we analyzed prostate cancer tissue samples. In this context miR-193a RNA was undetectable and MLL1 was highly expressed with concomitantly high levels of H3K4me, H3K4me2, and H3K4me3 enrichment in the promoters of MLL1 responsive genes. Finally, we showed that prolonged ectopic expression of miR-193a inhibits growth and cell migration, and induces apoptosis. Thus, while our study unveils amplitude of the epigenome, including miRnome it establishes that; (i) miR-193a directly target MLL1 mRNA, (ii) miR-193a impair MLL1 protein production, (iii) miR-193a reduces the overall methylation marks of the genome.
混合谱系白血病 1 (MLL1) 酶在调节与脊椎动物发育相关的基因方面发挥主要作用。细胞生理学和动态平衡由不同微环境中的 microRNAs 调节。在这项研究中,我们在 MLL1 基因转录本的 3'UTR 中鉴定了保守的 miR-193a 靶位点。利用野生型和突变型 3'UTR 构建体和荧光素酶报告基因检测,我们清楚地表明 miR-193a 直接靶向 MLL1 mRNA 的 3'UTR 区域。miR-193a 的异位表达调节了全局 H3K4 单、二和三甲基化水平,并影响了 CAV1 的表达,CAV1 是一种受 H3K4me3 特异性调节的基因。为了确定这种体外发现对异常生理条件的影响,我们分析了前列腺癌组织样本。在这种情况下,miR-193a RNA 检测不到,MLL1 高度表达,同时在 MLL1 反应基因的启动子中存在高水平的 H3K4me、H3K4me2 和 H3K4me3 富集。最后,我们表明,miR-193a 的长期异位表达抑制生长和细胞迁移,并诱导细胞凋亡。因此,虽然我们的研究揭示了表观基因组的幅度,包括 microRNA 组,但它确立了:(i)miR-193a 直接靶向 MLL1 mRNA,(ii)miR-193a 削弱 MLL1 蛋白的产生,(iii)miR-193a 降低基因组的整体甲基化标记。
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