Apoptosis repressor with caspase recruitment domain promotes cell proliferation and phenotypic modulation through 14-3-3ε/YAP signaling in vascular smooth muscle cells

J Mol Cell Cardiol. 2020 Oct:147:35-48. doi: 10.1016/j.yjmcc.2020.08.003. Epub 2020 Aug 6.

Abstract

Aims: In response to vascular injury, vascular smooth muscle cells (VSMC) may change from a contractile phenotype to a proliferative phenotype and consequently become conducive to neointima formation. Apoptosis repressor with caspase recruitment domain (ARC) was initially discovered as an endogenous apoptosis inhibitor, but whether ARC plays a role in VSMCs and whether it can participate in the regulation of atherosclerosis are unknown.

Methods and results: Protein and mRNA levels of ARC in tissues and cells were detected by western blot and quantitative real-time PCR. Immunofluorescence staining was used to detect the protein location, and immunohistochemistry was used to detect protein expression in tissues. VSMC proliferation was analysed using Cell Counting Kit-8 (CCK-8) and EdU assays, while migration was assessed by Transwell assay. Mechanistically, the direct binding between two proteins was verified by immunoprecipitation. We found that ARC expression was stimulated in VSMCs during cell proliferation. Our results also showed that ARC promoted cell proliferation and induced phenotypic modulation of VSMCs in vitro and vivo. Mechanistic studies demonstrated that ARC increased the nuclear localization of Yes associated protein (YAP) by binding to 14-3-3ε and that ARC played a role in promoting cell proliferation and phenotypic modulation. Additionally, the transcription factor p53 negatively regulated ARC expression at the transcriptional level during cell proliferation and phenotypic modulation.

Conclusions: Our findings define a novel role for ARC in the phenotypic transition of proliferating VSMCs, which may provide a new strategy for regulating neointimal formation.

Keywords: 14–3-3ε; ARC; Hippo-YAP signaling; Phenotypic modulation; VSMCs; p53.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 14-3-3 Proteins / metabolism*
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Animals
  • Apoptosis Regulatory Proteins / metabolism*
  • Cell Line
  • Cell Movement
  • Cell Proliferation
  • HEK293 Cells
  • Humans
  • Male
  • Mice, Inbred C57BL
  • Mitochondria / metabolism
  • Muscle Proteins / metabolism*
  • Muscle, Smooth, Vascular / cytology*
  • Myocytes, Smooth Muscle / metabolism*
  • Phenotype
  • Rats, Sprague-Dawley
  • Signal Transduction*
  • Transcription Factors / metabolism*
  • Transcription, Genetic
  • Tumor Suppressor Protein p53 / metabolism
  • Up-Regulation
  • YAP-Signaling Proteins

Substances

  • 14-3-3 Proteins
  • Adaptor Proteins, Signal Transducing
  • Apoptosis Regulatory Proteins
  • Muscle Proteins
  • NOL3 protein, human
  • Transcription Factors
  • Tumor Suppressor Protein p53
  • YAP-Signaling Proteins
  • YAP1 protein, human