MiR-197-3p affects angiogenesis and inflammation of endothelial cells by targeting CXCR2/COX2 axis

Decreased circulating miR-197-3p was found in patients with recurrent deep vein thrombosis (DVT), but the specific role of miR-197-3p needs further exploration. Materials and

MiR-197-3p affected viability, angiogenesis and inflammation of endothelial cells by targeting CXCR2/COX2 axis in vitro. Our findings provided a novel theoretical basis to investigate more effective therapies for DVT.

Venous blood samples were collected from DVT patients and healthy controls, and peripheral blood mononuclear cells (PBMCs) were isolated to examine the expression patterns of miR-197-3p, CXCR2 and COX2 by qRT-PCR. Human umbilical vein endothelial cells (HUVECs) were further used as a cellular model to investigate the role of the miR-197-3p/CXCR2/COX2 axis in regulating cell viability, angiogenesis, and inflammation, which were determined by MTT assay, Matrigel-based tube formation assay, and enzyme-linked immunosorbent assay, respectively. Dual-luciferase reporter assay was used to examine the interactions between miR-198-3p and CXCR2. Expression of NF-κB p65 was examined by western blot to investigate whether the NF-κB pathway was involved in the regulatory effect of miR-197-3p on DVT.

miR-197-3p was decreased in PBMCs of patients with DVT, while CXCR2 and COX2 were increased compared to the healthy controls. In HUVECs, overexpression of miR-197-3p reduced CXCR2 levels and inhibited cell viability, angiogenesis, and release of inflammatory cytokines including TNF-α, IL-1β, and IL-6, which were reversed by miR-197-3p inhibition. Dual-luciferase reporter assay indicated miR-197-3p directly bound to CXCR2. CXCR2 further upregulated the expression of COX2 and activated the NF-κB pathway, promoting cell viability, angiogenesis and release of inflammatory cytokines in HUVECs. The effect of miR-197-3p inhibition on cell viability, angiogenesis and inflammation of HUVECs could be reversed by CXCR2 silencing.