Knockdown of NEAT1 mitigates ox-LDL-induced injury in human umbilical vein endothelial cells via miR-30c-5p/TCF7 axis
Knockdown of NEAT1 mitigates ox-LDL-induced injury in human umbilical vein endothelial cells via miR-30c-5p/TCF7 axis
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DOI：
10.26355/eurrev_202009_23052
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Yu, H-B
中科院分区：
文献类型：
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作者：
Guo, J-T;Wang, L.;Yu, H-B
目的：动脉粥样硬化是一种炎症相关性疾病，对健康危害巨大。大量研究表明，长链非编码RNA（lncRNA）在动脉粥样硬化中发挥着重要作用，但核富集丰富转录本（NEAT 1）的分子机制尚未完全阐明。患者和方法：用氧化低密度脂蛋白（ox-LDL）处理人脐静脉内皮细胞（HUVEC），构建动脉粥样硬化模型。通过定量真实的时间-聚合酶链反应（qRT-PCR）检测NEAT 1、microRNA-30 c-5 p（miR-30 c-5 p）和转录因子7（TCF 7）的表达。MTT法检测细胞增殖活性，流式细胞仪检测细胞凋亡。采用Western blot法检测炎症相关蛋白的表达，酶联免疫吸附试验（ELISA）检测炎症细胞因子的浓度。结果：动脉粥样硬化患者血清及ox-LDL处理的HUVECs中NEAT 1表达上调。在ox-LDL处理的HUVECs中，NEAT 1的敲低促进增殖，但抑制凋亡和炎症。此外，NEAT 1靶向miR-30 c-5 p，并且miR-30 c-5 p的过表达逆转了HUVECs中ox-LDL诱导的效应。此外，miR-30 c-5 p直接抑制TCF 7的表达，NEAT 1通过上调miR-30 c-5 p抑制TCF 7的表达。结论：抑制NEAT 1基因表达可通过miR-30 c-5 p/TCF 7轴促进ox-LDL诱导的HUVECs增殖，并通过miR-30 c-5 p/TCF 7轴减轻细胞凋亡和炎症反应。NEAT 1加速了动脉粥样硬化治疗的进展，作为一个指示性元素。
OBJECTIVE: Atherosclerosis is an inflammation-associated disease resulting in a huge health hazard. Abundance of researches showed that long non-coding RNAs (lncRNAs) played vital roles in atherosclerosis, but the molecular mechanism of nuclear-enriched abundant transcript (NEAT1) has not been fully elucidated yet.PATIENTS AND METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with oxidized low-density lipoprotein (ox-LDL) for constructing the model of atherosclerosis. The detection of NEAT1, microRNA-30c-5p (miR-30c-5p), and transcription factor 7 (TCF7) expression was implemented by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Cell proliferation and apoptosis were measured by 3-(4, 5-dimethylthiazol-2-y1)-2, 5-diphenyl tetrazolium bromide (MTT) and flow cytometry, respectively. The levels of apoptosis-associated proteins were examined through Western blot and the concentrations of inflammatory cytokines were determined by enzyme-linked immunosorbent assay (ELISA). The targeted relationship was analyzed by Dual-Luciferase reporter assay.RESULTS: NEAT1 was upregulated in serum of patients with atherosclerosis and HUVECs treated with ox-LDL. Knockdown of NEAT1 exerted the promotion of proliferation but suppression of apoptosis and inflammation in ox-LDL-treated HUVECs. Moreover, NEAT1 targeted miR-30c-5p and the overexpression of miR-30c-5p reversed the ox-LDL-induced effects in HUVECs. Furthermore, miR-30c-5p directly refrained the TCF7 level, and NEAT1 repression decreased the expression of TCF7 by upregulating miR-30c-5p. The knockdown of NEAT1 afforded the protective effect for HUVECs treated with ox-LDL through miR-30c-5p/TCF7 axis.CONCLUSIONS: The knockdown of NEAT1 overtly motivated proliferation but alleviated the apoptosis and inflammation in ox-LDL-treated HUVECs by miR-30c-5p/TCF7 axis. NEAT1 accelerated the progression of atherosclerosis therapies, functioning as an indicative element.