Department of Vascular Surgery, Taizhou University Affiliated Municipal Hospital.
Int Heart J. 2023;64(2):252-262. doi: 10.1536/ihj.22-148.
Atherosclerosis (AS) is the main reason for most cardiovascular diseases. Circular RNA hsa_circ_0044073 (circ_0044073) has been found to promote AS progression. However, the specific regulatory mechanism of circ_0044073 in AS progression remains unclear.In this study, oxidized low-density lipoprotein (Ox-LDL) -stimulated human vascular smooth muscle cells (VSMCs) were used as AS cell models. The expression changes of circ_0044073 in serum samples and Ox-LDL-stimulated human VSMCs were assessed via real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, proliferation, colony formation, migration, and invasion were assessed using 3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide (MTT), 5-ethynyl-2'-deoxyuridine (EDU), colony formation, and transwell assays. Some protein levels were detected via Western blotting. The regulatory mechanism of circ_0044073 was predicted using bioinformatics analysis and validated by dual-luciferase reporter and RNA pull-down assays.We observed an overt increase in circ_0044073 expression in serum samples derived from AS patients and Ox-LDL-stimulated human VSMCs. Circ_0044073 was identified as a miR-377-3p sponge. Either circ_0044073 knockdown or miR-377-3p overexpression could impair Ox-LDL-induced human VSMC proliferation, migration, invasion, and inflammation. AURKA served as a miR-377-3p target, and circ_0044073 regulated AURKA expression by adsorbing miR-377-3p. Furthermore, AURKA overexpression partly reversed the effects of circ_0044073 inhibition on Ox-LDL-induced human VSMC proliferation, migration, invasion, and inflammation.Circ_0044073 promoted AS progression by elevating AURKA expression by functioning as a miR-377-3p sponge. Providing a proof-of-concept demonstration to support circ_0044073 might be a target for AS treatment.
动脉粥样硬化(AS)是大多数心血管疾病的主要原因。已经发现环状 RNA hsa_circ_0044073(circ_0044073)可促进 AS 进展。然而,circ_0044073 在 AS 进展中的具体调节机制仍不清楚。在这项研究中,使用氧化低密度脂蛋白(Ox-LDL)刺激的人血管平滑肌细胞(VSMCs)作为 AS 细胞模型。通过实时定量聚合酶链反应(RT-qPCR)评估血清样本和 Ox-LDL 刺激的人 VSMCs 中 circ_0044073 的表达变化。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)、5-乙炔基-2'-脱氧尿苷(EDU)、集落形成和 Transwell 测定评估细胞活力、增殖、集落形成、迁移和侵袭。通过 Western blot 检测某些蛋白水平。通过生物信息学分析预测 circ_0044073 的调节机制,并通过双荧光素酶报告和 RNA 下拉测定进行验证。我们观察到 AS 患者血清样本和 Ox-LDL 刺激的人 VSMCs 中 circ_0044073 的表达明显增加。circ_0044073 被鉴定为 miR-377-3p 的海绵。circ_0044073 敲低或 miR-377-3p 过表达均可损害 Ox-LDL 诱导的人 VSMC 增殖、迁移、侵袭和炎症。AURKA 是 miR-377-3p 的靶标,circ_0044073 通过吸附 miR-377-3p 调节 AURKA 表达。此外,AURKA 过表达部分逆转了 circ_0044073 抑制对 Ox-LDL 诱导的人 VSMC 增殖、迁移、侵袭和炎症的影响。Circ_0044073 通过作为 miR-377-3p 海绵来升高 AURKA 表达,从而促进 AS 进展。为支持 circ_0044073 可能成为 AS 治疗的靶点提供了概念验证证据。