Hsa-circ_0010283 Regulates Oxidized Low-Density Lipoprotein-Induced Proliferation and Migration of Vascular Smooth Muscle Cells by Targeting the miR-133a-3p/Pregnancy-Associated Plasma Protein A Axis

Circ J. 2020 Nov 25;84(12):2259-2269. doi: 10.1253/circj.CJ-20-0345. Epub 2020 Nov 6.

Abstract

Background: The dysfunction of vascular smooth muscle cells (VSMCs) contributes to the development of atherosclerosis. This study aimed to investigate the role of circular RNA-0010283 (circ_0010283) in oxidized low-density lipoprotein (ox-LDL)-treated VSMCs and the associated action mechanism.

Methods and results: The expression of circ_0010283 was investigated using quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was monitored by using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cell apoptosis was detected by using flow cytometry assay. A transwell assay was performed to observe migration and invasion, and a scratch assay was implemented to test migration. The expression of proliferation, apoptosis and migration/invasion-related proteins was measured by using a western blot. The targeted relationship was predicted by using a bioinformatics tool (Starbase) and verified by using a dual-luciferase reporter assay, a RNA immunoprecipitation (RIP) assay and a RNA pull-down assay. circ_0010283 was highly expressed in serum samples from atherosclerosis patients and ox-LDL-treated human VSMCs (HVSMCs). circ_0010283 knockdown suppressed ox-LDL-induced proliferation, migration and invasion in HVSMCs. MicroRNA-133a-3p (miR-133a-3p) was confirmed as a target of circ_0010283, and miR-133a-3p deficiency reversed the effects of circ_0010283 knockdown. Moreover, pregnancy-associated plasma protein A (PAPPA) was targeted by miR-133a-3p, and PAPPA overexpression reversed the effects of miR-133a-3p restoration. Interestingly, circ_0010283 could regulate PAPPA expression by mediating miR-133a-3p.

Conclusions: circ_0010283 participated in ox-LDL-induced dysfunctions of HVSMCs by modulating the miR-133a-3p/PAPPA pathway, suggesting that circ_0010283 might be associated with atherosclerosis pathogenesis.

Keywords: Atherosclerosis; Circular RNA-0010283 (circ_0010283); MicroRNA-133a-3p (miR-133a-3p); Pregnancy-associated plasma protein A (PAPPA); Vascular smooth muscle cells (VSMCs).

MeSH terms

  • Apoptosis
  • Atherosclerosis* / genetics
  • Blood Proteins
  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Humans
  • Lipoproteins, LDL / metabolism*
  • MicroRNAs* / genetics
  • Muscle, Smooth, Vascular / cytology
  • Myocytes, Smooth Muscle / cytology*
  • Pregnancy-Associated Plasma Protein-A / genetics*
  • RNA, Circular* / genetics

Substances

  • Blood Proteins
  • Lipoproteins, LDL
  • MIRN133 microRNA, human
  • MicroRNAs
  • RNA, Circular
  • oxidized low density lipoprotein
  • Pregnancy-Associated Plasma Protein-A
  • PAPPA protein, human