A regulatory circuit involving miR-143 and DNMT3a mediates vascular smooth muscle cell proliferation induced by homocysteine.
A regulatory circuit involving miR-143 and DNMT3a mediates vascular smooth muscle cell proliferation induced by homocysteine.
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DOI：
10.3892/mmr.2015.4558
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发表时间：
2016
影响因子：
3.4
通讯作者：
Huiping Zhang;Yanhua Wang;Cheng-jian Cao;Xiaoming Yang;Sheng-chao Ma;Xue-Bo Han;Xiaoling Yang;A. Yang;Jue Tian;Hua Xu;Minghao Zhang;Yideng Jiang
中科院分区：
文献类型：
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作者：
Huiping Zhang;Yanhua Wang;Cheng-jian Cao;Xiaoming Yang;Sheng-chao Ma;Xue-Bo Han;Xiaoling Yang;A. Yang;Jue Tian;Hua Xu;Minghao Zhang;Yideng Jiang
越来越多的证据表明，同型半胱氨酸（Hcy）是动脉粥样硬化（AS）的独立危险因素。同型半胱氨酸可促进血管平滑肌细胞（VSMC）增殖，在AS的发病和发展中起关键作用。本研究旨在探讨Hcy诱导microRNA（miR）-143介导VSMCs增殖的表观遗传学调控机制。3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide检测结果显示，与对照组相比，100 mM Hcy处理后VSMC增殖增加了1.39倍。使用逆转录-定量聚合酶链反应分析确定，与对照组相比，Hcy组中miR-143的水平显著下调。此外，与对照组相比，Hcy处理的VSMCs中miR-143甲基化水平显著增加，并在用DNA甲基转移酶（DNMT）3a小干扰RNA转染后降低，使用甲基化特异性PCR测定。在分别转染前miR-143和miR-143抑制剂的VSMC中，DNMT 3a荧光素酶的活性也相应地改变。此外，在VSMC中观察到miR-143的表达与DNMT 3的mRNA和蛋白表达呈负相关。总而言之，这些发现表明DNMT 3a是miR-143的直接靶点，并且DNMT 3的上调是Hcy诱导的VSMC增殖中miR-143高甲基化的原因。
Accumulating evidence has suggested that homocysteine (Hcy) is an independent risk factor for atherosclerosis (AS). Hcy can promote vascular smooth muscle cell (VSMC) proliferation, which is pivotal in the pathogenesis and progression of AS. The aim of the present study was to investigate the epigenetic regulatory mechanism of microRNA (miR)‑143‑mediated VSMCs proliferation induced by Hcy. The results of a 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphe‑nyltetrazolium bromide assay revealed that VSMC proliferation was increased by 1.39‑fold following treatment with 100 mM Hcy, compared with the control group. The levels of miR‑143 were markedly downregulated in the Hcy group, compared with the control group, as determined using reverse transcription‑quantitative polymerase chain reaction analysis. In addition, the level of miR‑143 methylation was increased markedly in the VSMCs treated with Hcy, compared with the control, and was reduced following transfection with DNA methyltransferase (DNMT)3a small interfering RNA, determined using methylation‑specific‑PCR. The activities of DNMT3a luciferase were also altered accordingly in VSMCs transfected with pre‑miR‑143 and miR‑143 inhibitor, respectively. In addition, the expression of miR‑143 was observed to be inversely correlated with the mRNA and protein expression of DNMT3 in the VSMCs. Taken together, these findings suggest that DNMT3a is a direct target of miR‑143, and that the upregulation of DNMT3 is responsible for the hypermethylation of miR‑143 in Hcy-induced VSMC proliferation.