Circ_0020397 regulates the viability of vascular smooth muscle cells by up-regulating GREM1 expression via miR-502-5p in intracranial aneurysm

Abstract

AimsCirc_0020397 has been found to be down-regulated in intracranial aneurysm (IA), and deregulation of circ_0020397 involved in the regulation of vascular smooth muscle cells (VSMCs) proliferation. However, the mechanism by which circ_0020397 implicates in VSMC dysfunction in IA remains vague. Materials and methodsThe expression of circ_0020397, miR-502-5p and Gremlin 1 (GREM1) was detected using quantitative real-time polymerase chain reaction. Cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Protein levels of proliferating cell nuclear antigen (PCNA) and GREM1 were measured using western blot. The interaction between miR-502-5p and circ_0020397 or GREM1 was confirmed by dual-luciferase reporter, RNA pull-down and RNA immunoprecipitation assay. Key findingsCirc_0020397 or GREM1 expression was decreased in VSMCs isolated from IA patients, and overexpression of circ_0020397 or GREM1 promoted VSMC viability and elevated PCNA expression level, while inhibition of them showed opposite effects. MiR-502-5p was confirmed to directly bind to circ_0020397 or GREM1, and miR-502-5p reversed the effects of circ_0020397 on VSMC viability and PCNA level. Besides, miR-502-5p overexpression suppressed VSMC viability and reduced PCNA level, while these effects were attenuated by GREM1 up-regulation. Importantly, circ_0020397 could regulate GREM1 expression via miR-502-5p in VSMCs. SignificanceCirc_0020397 played an important role in phenotypic modulation in IA by promoting VSMC viability via miR-502-5p/GREM1 axis, suggesting a novel insight into IA pathogenesis and new targets for IA molecular therapy.