Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, China.
Liaoning Provincial Core Lab of Medical Molecular Biology, Dalian Medical University, Dalian, China.
J Physiol Biochem. 2021 Feb;77(1):47-61. doi: 10.1007/s13105-020-00779-5. Epub 2021 Jan 7.
Endothelial injury, which can cause endothelial inflammation and dysfunction, is an important mechanism for the development of atherosclerotic plaque. This study aims to investigate the functional role of miR-520c-3p in vascular endothelium during inflammatory diseases such as atherosclerosis. Quantitative real-time PCR was used to detect miR-520c-3p expression in in human umbilical vein endothelial cells (HUVECs) after treatment with platelet-derived growth factor (PDGF). Furthermore, the effects of miR-520c-3p overexpression and silencing on cell proliferation, adhesion, and apoptosis were assessed. Bioinformatics analysis and Biotin-labeled miRNA pull-down assay were used to confirm the targets of miR-520-3p. Then, the effects of miR-520c-3p on AKT and NF-κB signaling pathways were detected by western blot. Herein, we observed that the expression level of miR-520c-3p was downregulated in HUVECs under PDGF stimulation. Overexpression of miR-520c-3p not only decreased cell adhesion but also promoted proliferation and inhibited apoptosis to protect the viability of endothelial cells. It was confirmed that RELA is the target of miR-520c-3p. MiR-520c-3p inhibited the protein phosphorylation of AKT and RELA, and si-RELA reversed the promotion of AKT and RELA protein phosphorylation by anti-miR-520c-3p. In summary, our study suggested that miRNA-520c-3p targeting RELA through AKT and NF-κB signaling pathways regulated the proliferation, apoptosis, and adhesion of vascular endothelial cells. We conclude that miR-520c-3p may play an important role in the suppression of endothelial injury, which could serve as a biomarker and therapeutic target for atherosclerosis.
内皮损伤可导致内皮炎症和功能障碍,是动脉粥样硬化等炎症性疾病中粥样斑块发展的重要机制。本研究旨在探讨 miR-520c-3p 在血管内皮细胞炎症性疾病中的功能作用。采用血小板衍生生长因子(PDGF)处理人脐静脉内皮细胞(HUVEC)后,用实时定量 PCR 检测 miR-520c-3p 的表达。此外,还评估了 miR-520c-3p 过表达和沉默对细胞增殖、黏附和凋亡的影响。生物信息学分析和生物素标记 miRNA 下拉实验用于证实 miR-520c-3p 的靶标。然后,通过 Western blot 检测 miR-520c-3p 对 AKT 和 NF-κB 信号通路的影响。结果显示,PDGF 刺激下 HUVECs 中 miR-520c-3p 的表达水平下调。miR-520c-3p 过表达不仅降低了细胞黏附,还促进了增殖,抑制了凋亡,从而保护了内皮细胞的活力。证实 RELA 是 miR-520c-3p 的靶标。miR-520c-3p 抑制 AKT 和 RELA 蛋白磷酸化,si-RELA 逆转了抗 miR-520c-3p 对 AKT 和 RELA 蛋白磷酸化的促进作用。综上所述,本研究表明,miR-520c-3p 通过 AKT 和 NF-κB 信号通路靶向 RELA,调节血管内皮细胞的增殖、凋亡和黏附。我们得出结论,miR-520c-3p 可能在抑制内皮损伤中发挥重要作用,可作为动脉粥样硬化的生物标志物和治疗靶点。