MicroRNA-99a-5p alleviates atherosclerosis via regulating Homeobox A1
MicroRNA-99a-5p alleviates atherosclerosis via regulating Homeobox A1
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DOI：
10.1016/j.lfs.2019.116664
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发表时间：
2019-09-01
期刊：
影响因子：
6.1
通讯作者：
Jing, Bao
中科院分区：
文献类型：
--
作者：
Han, Zhiyang;Guan, Yinghui;Jing, Bao
目的：已证实microRNAs参与动脉粥样硬化的发生发展。本研究旨在探讨miR-99a-5p及其靶基因同源盒A1(Homeobox A1，HOXA1)在动脉粥样硬化中的作用。用TargetScan和miRDB对microRNAs的靶基因进行预测。荧光素酶报告实验证实miR-99a-5p与HOXA1结合。在体内实验中，建立了高脂饮食诱导的载脂蛋白E基因敲除小鼠动脉粥样硬化模型。苏木精-伊红(H&E)、油红O和Masson三色染色检测动脉粥样硬化病变。实时定量聚合酶链式反应检测miR-99a-5p和HOXA1基因的表达水平。通过免疫印迹和免疫组织化学分析检测HOXA1和迁移相关蛋白的表达。关键发现：miR-99a-5p通过靶向HOXA1 mRNA的3‘UTR而抑制HOXA1的表达。增强的HOXA1显著促进ASMC的增殖、迁移和侵袭。此外，miR-99a-5p的过表达抑制了HOXA1刺激的ASMC的增殖、迁移和侵袭，而miR99a-5p的抑制则逆转了HOXA1基因敲除对ASMC这些行为的影响。在体内，miR-99a-5p的特异性过表达显著减轻动脉粥样硬化形成的病变，同时HOXA1的mRNA和蛋白表达水平显著下调。意义：我们首次证明miR-99a-5p可能是一种潜在的动脉粥样硬化抑制剂，miR-99a-5p部分通过靶向HOXA1发挥作用。
Aims: MicroRNAs have been demonstrated to be involved in the development of atherosclerosis. The present study aimed to evaluate the effect of miR-99a-5p and its target gene Homeobox A1 (HOXA1) in atherosclerosis.Main methods: The biological functions of miR-99a-5p on human aortic smooth muscle cells (ASMCs) were assessed by MTT, wound healing and transwell assays. The target genes of microRNAs were predicted by TargetScan and miRDB. The binding of miR-99a-5p and HOXA1 was confirmed by luciferase reporter assay. In the in vivo study, high-fat diet-induced atherosclerosis model was established in Apolipoprotein E knockout mice. Hematoxylin-eosin (H&E), oil Red O and Masson trichrome staining were performed for determination of atherosclerotic lesion. The levels of miR-99a-5p and HOXA1 mRNA were detected by real-time PCR. HOXA1 and migration-associated protein levels were detected by western blot or immunohistochemistry analysis.Key findings: MiR-99a-5p inhibited HOXA1 expression by targeting 3'UTR of HOXA1 mRNA. Enforced HOXA1 significantly promoted the proliferation, migration, and invasion of ASMCs. Furthermore, miR-99a-5p overexpression inhibited the proliferation, migration, and invasion of ASMCs stimulated by HOXA1, whereas miR99a-5p inhibition reversed the effects of HOXA1 knockdown on these behaviours of ASMCs. In vivo, the specific overexpression of miR-99a-5p significantly abated atherosclerotic lesions formatted, accompanied with a significant down-regulation of HOXA1 mRNA and protein expression levels.Significance: We demonstrate for first time that miR-99a-5p may serve as a potential inhibitor of the atherosclerosis, and miR-99a-5p plays its role partially through targeting HOXA1.