CircTM7SF3 contributes to oxidized low-density lipoprotein-induced apoptosis, inflammation and oxidative stress through targeting miR-206/ASPH axis in atherosclerosis cell model in vitro

BMC Cardiovasc Disord. 2021 Feb 2;21(1):51. doi: 10.1186/s12872-020-01800-x.

Abstract

Background: Atherosclerosis (AS) is a chronic inflammatory disorder. The aim of our study was to explore the role of circular RNA (circRNA) transmembrane 7 superfamily member 3 (circTM7SF3) in AS progression.

Methods: Experiments were conducted using oxidized low-density lipoprotein (ox-LDL)-induced THP-1-derived macrophages and differentiated human monocyte-derived macrophages (hMDMs). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circTM7SF3, its linear form TM7SF3, microRNA-206 (miR-206) and aspartyl (asparaginyl) β-hydroxylase (ASPH) messenger RNA (mRNA). Cell viability and apoptosis were examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Cell inflammation was analyzed by measuring the production of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) using enzyme-linked immunosorbent assay (ELISA) kits. Cell oxidative stress was assessed through analyzing the levels of oxidative stress markers using their corresponding commercial kits. Dual-luciferase reporter assay and RNA-pull down assay were used to confirm the interaction between miR-206 and circTM7SF3 or ASPH. The protein level of ASPH was examined by Western blot assay.

Results: CircTM7SF3 level was markedly increased in the serum samples of AS patients and ox-LDL-induced THP-1-derived macrophages compared with their matching counterparts. ox-LDL induced-damage in THP-1 cells was partly attenuated by the interference of circTM7SF3. MiR-206 was a downstream molecular target of circTM7SF3. Si-circTM7SF3-mediated effects in ox-LDL-induced THP-1-derived macrophages were partly ameliorated by the addition of anti-miR-206. MiR-206 directly interacted with ASPH mRNA. CircTM7SF3 silencing reduced the expression of ASPH partly through up-regulating miR-206 in THP-1-derived macrophages. ASPH overexpression partly counteracted the effects induced by miR-206 overexpression in ox-LDL-induced THP-1-derived macrophages.

Conclusion: CircTM7SF3 contributed to ox-LDL-induced injury in AS cell model through up-regulating the expression of ASPH via targeting miR-206.

Keywords: ASPH; Atherosclerosis; Ox-LDL; circTM7SF3; miR-206.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Apoptosis / drug effects*
  • Atherosclerosis / enzymology*
  • Atherosclerosis / genetics
  • Atherosclerosis / pathology
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism*
  • Case-Control Studies
  • Female
  • Gene Expression Regulation
  • Humans
  • Inflammation Mediators / metabolism*
  • Interleukin-6 / metabolism
  • Lipoproteins, LDL / toxicity*
  • Macrophages / drug effects*
  • Macrophages / enzymology
  • Macrophages / pathology
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Middle Aged
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism*
  • Muscle Proteins / genetics
  • Muscle Proteins / metabolism*
  • Oxidative Stress / drug effects*
  • RNA, Circular / genetics
  • RNA, Circular / metabolism*
  • Signal Transduction
  • THP-1 Cells
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Calcium-Binding Proteins
  • IL6 protein, human
  • Inflammation Mediators
  • Interleukin-6
  • Lipoproteins, LDL
  • MIRN206 microRNA, human
  • Membrane Proteins
  • MicroRNAs
  • Muscle Proteins
  • RNA, Circular
  • TNF protein, human
  • Tumor Necrosis Factor-alpha
  • oxidized low density lipoprotein
  • Mixed Function Oxygenases
  • ASPH protein, human