Dartpartment of Cardiovascular Surgery, The Third People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang, China.
J Cardiovasc Pharmacol. 2021 May 1;77(5):603-613. doi: 10.1097/FJC.0000000000000983.
Circular RNAs have shown regulatory functions in atherosclerosis (AS) progression. Here, we explored the role and working mechanism of circ_0000345 in the AS cell model in vitro. Quantitative real-time polymerase chain reaction was applied to measure the enrichment of circ_0000345, microRNA-129-5p (miR-129-5p), and ten-eleven translocation-2 (TET2) messenger RNA. Cell Counting Kit 8 assay was used to analyze cell viability of human umbilical vein endothelial cells (HUVECs). Flow cytometry was conducted to assess cell apoptosis and cell cycle progression. The target relationship between miR-129-5p and circ_0000345 or TET2 was verified by the dual-luciferase reporter assay. The Western blot assay was used to analyze the protein level of TET2. Circ_0000345 abundance was reduced in serum samples of AS patients and AS cell model compared with their matching counterparts. Circ_0000345 overexpression promoted cell viability and cell cycle progression and hampered cell apoptosis in HUVECs induced by oxidized low-density lipoprotein. MiR-129-5p was a target of circ_0000345 and circ_0000345 attenuated ox-LDL-induced damage in HUVECs through sponging miR-129-5p. MiR-129-5p bound to the 3' untranslated region (3'UTR) of TET2, and miR-129-5p functioned in ox-LDL-induced HUVECs by targeting TET2. Circ_0000345 enhanced TET2 messenger RNA and protein expression through sponging miR-129-5p in HUVECs. Circ_0000345 attenuated ox-LDL-mediated injury in HUVECs through targeting miR-129-5p/TET2 axis. Increasing the levels of circ_0000345 and TET2 might be a novel insight into AS treatment.
环状 RNA 在动脉粥样硬化（AS）进展中表现出调节功能。在这里，我们研究了 circ_0000345 在体外 AS 细胞模型中的作用和工作机制。实时定量聚合酶链反应用于测量 circ_0000345、microRNA-129-5p（miR-129-5p）和 ten-eleven 易位-2（TET2）信使 RNA 的丰度。细胞计数试剂盒 8 用于分析人脐静脉内皮细胞（HUVEC）的细胞活力。流式细胞术用于评估细胞凋亡和细胞周期进程。通过双荧光素酶报告基因检测验证 miR-129-5p 与 circ_0000345 或 TET2 的靶关系。Western blot 检测用于分析 TET2 蛋白水平。与相应的对照相比，AS 患者的血清样本和 AS 细胞模型中的 circ_0000345 丰度降低。circ_0000345 过表达促进了氧化型低密度脂蛋白诱导的 HUVEC 细胞活力和细胞周期进程，并抑制了细胞凋亡。miR-129-5p 是 circ_0000345 的靶标，circ_0000345 通过海绵 miR-129-5p 减轻了 ox-LDL 诱导的 HUVEC 损伤。miR-129-5p 结合到 TET2 的 3'非翻译区（3'UTR），miR-129-5p 通过靶向 TET2 在 ox-LDL 诱导的 HUVEC 中发挥作用。circ_0000345 通过海绵 miR-129-5p 增强了 HUVEC 中的 TET2 信使 RNA 和蛋白表达。circ_0000345 通过靶向 miR-129-5p/TET2 轴减轻了 ox-LDL 介导的 HUVEC 损伤。增加 circ_0000345 和 TET2 的水平可能为 AS 治疗提供新的思路。
