The Studies On The Role Of MiR-205 In Angiogenesis Of Endothelial Progenitor Cell And Deep Vein Thrombosis Resolution

Objective:To clarify the role of miR-205 in the cell proliferation,apoptosis,migration and angiogenesis of endothelial progenitor cells(EPCs);To test the effects of miR-205 on the homing ability of EPCs to deep vein thrombosis sites and its value in the treatment of deep vein thrombosis;To investigate the target of miR-205 and its underlying mechanism of regulating angiogenesis in EPCs.Methods:1.EPCs were divided into four groups:N group(untreated EPCs),NC group(LV3-NC vector plasmid-transfected EPCs),miR-205 mimics group(miR-205 overexpression group),miR-205 inhibitor group(miR-205 down-regulation group).Fluorescence microscopy and RT-qPCR were performed to detect the infection efficiency of lentiviruses.2.The migration rate,invasion and angiogenesis of EPCs were detected by wound healing assay,transwell assay and tube formation assay,respectively;The cell proliferation was quantified by CCK8 assay;Cell apoptosis was detected by flow cytometry;In order to test the role of miR-205 in the regulation of angiogenesis in vivo,EPCs were mixed with Matrigel plugs and injected into nude mice to simulate angiogenesis in vivo.HE staining was used to observe the formation of new blood vessels.3.A model of jugular vein thrombosis in nude mice was established,and EPCs overexpressing miR-205 with green fluorescent protein(GFP-miR-205 EPCs)were transplanted into the thrombosis.One week later,the jugular vein with thrombus was removed,and the size and weight were performed to demonstrate deep vein thrombosis resolution and recanalization;The number of EPCs homing to the deep vein thrombosis sites was measured by fluorescence analysis.4.The target of miR-205 and its mechanism of regulating EPCs were investigated via dual luciferase assay,RT-qPCR assay,western blot assay and co-transfection to test the cell function.Results:1.The results of fluorescence and RT-qPCR showed that lentiviral infection was successful.2.Overexpression of miR-205 significantly enhanced the migration,invasion,tube formation and proliferation of EPCs and inhibited cell apoptosis,which could significantly enhance the ability of angiogenesis in vivo;On the contrary,miR-205 observably suppressed the migration,invasion,angiogenesis and proliferative capacity of EPCs and promoted apoptosis.3.miR-205 enhanced the homing ability of EPCs to deep vein thrombosis sites and promoted thrombosis dissolution and recanalization.4.The target of miR-205 was PTEN in EPCs.Conclusion:MiR-205 enhanced EPC proliferation,homing ability,angiogenesis in vitro and in vivo,and inhibited cell apoptosis via targeting PTEN,and it facilitated the deep vein thrombosis dissolution and recanalization.