Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, PR China.
Department of Emergency Internal Medicine, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, PR China.
J Cell Biochem. 2019 Sep;120(9):14670-14678. doi: 10.1002/jcb.28728. Epub 2019 Apr 23.
Long noncoding RNAs (lncRNAs) play important roles in endothelium development. A lncRNA, LEF1-AS1, is recently emerging as a potent mediator of the proliferation and migration of a number of cells, including smooth muscle cells. However, the effects of LEF1-AS1 in atherosclerosis remains largely unknown. Specimens from patients with coronary artery atherosclerosis were collected. The quantitative real-time polymerase chain reaction was used to analyze levels of LEF1-AS1 and microRNA-544a (miR-544a). Western blot analysis was used to assess PTEN, P-Akt, and T-Akt protein expression. Proliferation, migration, and invasion of cells were analyzed by cell counting kit-8 assay, scratch wound assay, and transwell assay, respectively. The interaction between LEF1-AS1, miR-544a, and PTEN was probed using bioinformatical analysis and dual-luciferase assay. In plasma and tissue of patients with coronary artery atherosclerosis, LEF1-AS1 was upregulated and miR-544a was downregulated. A negative correlation was found between LEF1-AS1 and miR-544a. miR-544a overexpression reversed the inhibition of LEF1-AS1 in smooth muscle cell proliferation and invasion, which were mediated through the PTEN pathway. LEF1-AS1 regulates smooth muscle cell proliferation and migration through the miR-544a/PTEN axis, indicating that LEF1-AS1 may be a potential therapeutic target in atherosclerosis.
长链非编码 RNA(lncRNA)在血管内皮细胞发育中发挥重要作用。长链非编码 RNA,LEF1-AS1,作为许多细胞,包括平滑肌细胞增殖和迁移的有效介质,最近逐渐受到关注。然而,LEF1-AS1 在动脉粥样硬化中的作用仍然知之甚少。收集了患有冠状动脉粥样硬化的患者的标本。采用定量实时聚合酶链反应分析 LEF1-AS1 和 microRNA-544a(miR-544a)的水平。Western blot 分析用于评估 PTEN、P-Akt 和 T-Akt 蛋白表达。通过细胞计数试剂盒-8 测定、划痕愈合测定和 Transwell 测定分别分析细胞增殖、迁移和侵袭。通过生物信息学分析和双荧光素酶报告基因检测探究 LEF1-AS1、miR-544a 和 PTEN 之间的相互作用。在患有冠状动脉粥样硬化的患者的血浆和组织中,LEF1-AS1 上调,miR-544a 下调。发现 LEF1-AS1 与 miR-544a 之间呈负相关。miR-544a 的过表达逆转了 LEF1-AS1 对平滑肌细胞增殖和侵袭的抑制作用,该作用是通过 PTEN 通路介导的。LEF1-AS1 通过 miR-544a/PTEN 轴调节平滑肌细胞的增殖和迁移,表明 LEF1-AS1 可能是动脉粥样硬化的潜在治疗靶点。