Department of Cardiology, Yiwu Central Hospital, Yiwu 322000, China.
Acta Biochim Biophys Sin (Shanghai). 2020 Sep 8;52(9):967-974. doi: 10.1093/abbs/gmaa087.
Studies have shown that long non-coding RNAs (lncRNA) play critical roles in coronary atherosclerotic heart disease (CAD). However, the function of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in CAD is unclear. In this study, we aimed to investigate the functions of lncRNA NEAT1 in CAD. RT-PCR and western blot analysis were carried out to examine the expressions of related RNAs. Colony formation assay, cell proliferation assay, apoptosis assay, and dual-luciferase reporter assay were conducted to investigate the abilities of colony migration, cell proliferation, apoptosis, and targeting. The results showed that NEAT1 was up-regulated in CAD blood samples and in human coronary endothelial cells (HCAECs). Transfection of pcNEAT1 significantly inhibited the survival rate of HCAECs and induced apoptosis of HCAECs. MiR-140-3p was down-regulated in HCAECs. NEAT1 directly targeted miR-140-3p, and the expression of miR-140-3p was inversely correlated with the expression of NEAT1 in CAD patients. In addition, co-transfection of NEAT1 with miR-140-3p mimic reversed the effect of pcNEAT1 on cell viability and apoptosis. mitogen-activated protein kinase 1 (MAPK1) was proved to be a target gene of miR-140-3p, and the miR-140-3p mimic was shown to reduce the expression of MAPK1 in HCAECs. pcNEAT1 significantly increased the expression level of MAPK1, while shNEAT1 significantly reduced the expression level of MAPK1. Our results revealed that lncRNA NEAT1 increased cell viability and inhibited CAD cell apoptosis possibly by activating the miR-140-3p/MAPK1 pathway, and lncRNA NEAT1 might serve as a potential therapeutic target for CAD.
研究表明，长链非编码 RNA（lncRNA）在冠状动脉粥样硬化性心脏病（CAD）中发挥着关键作用。然而，lncRNA 核富集丰富转录本 1（NEAT1）在 CAD 中的功能尚不清楚。在本研究中，我们旨在研究 lncRNA NEAT1 在 CAD 中的作用。通过 RT-PCR 和 Western blot 分析检测相关 RNA 的表达。通过集落形成试验、细胞增殖试验、凋亡试验和双荧光素酶报告基因试验检测细胞集落迁移、细胞增殖、凋亡和靶向能力。结果表明，CAD 血液样本和人冠状动脉内皮细胞（HCAEC）中 NEAT1 表达上调。pcNEAT1 的转染显著抑制了 HCAEC 的存活率并诱导了 HCAEC 的凋亡。miR-140-3p 在 HCAEC 中下调。NEAT1 直接靶向 miR-140-3p，CAD 患者中 NEAT1 的表达与 miR-140-3p 的表达呈负相关。此外，NEAT1 与 miR-140-3p 模拟物共转染逆转了 pcNEAT1 对细胞活力和凋亡的影响。丝裂原活化蛋白激酶 1（MAPK1）被证明是 miR-140-3p 的靶基因，miR-140-3p 模拟物显示降低了 HCAEC 中 MAPK1 的表达。pcNEAT1 显著增加了 MAPK1 的表达水平，而 shNEAT1 则显著降低了 MAPK1 的表达水平。我们的研究结果表明，lncRNA NEAT1 通过激活 miR-140-3p/MAPK1 通路增加细胞活力并抑制 CAD 细胞凋亡，lncRNA NEAT1 可能成为 CAD 的潜在治疗靶点。
