MicroRNA-195 regulates proliferation, migration, angiogenesis and autophagy of endothelial progenitor cells by targeting GABARAPL1.

Jianwen Mo,Renze Yang,Daifen Zhang

doi:10.1042/bsr20160139

Abstract

Deep vein thrombosis (DVT) is a common type of venous thrombosis. Successful resolution of DVT-related thrombi is important in the treatment of DVT. Endothelial progenitor cells (EPCs) have emerged as a promising therapeutic choice for DVT-related thrombus resolution; however, the clinical application of EPCs faces many challenges. In the present study, the expression of miR-582, miR-195 and miR-532 under hypoxic or normoxic conditions was measured using quantitative real-time PCR analysis (qRT-PCR) and the results showed that the increased fold of miR-195 was highest in human EPCs (hEPCs) under hypoxic conditions. Then the role and regulating mechanism of miR-195 in improving the function of EPCs was investigated. To investigate the effect of miR-195 inhibition on the autophagy of hEPCs, the expression of the autophagy-related genes LC3B and beclin1 was examined using western blotting, and the formation of autophagosomes was observed using TEM. The results indicated that the inhibition of miR-195 expression could promote autophagy of hEPCs. In addition, we investigated the role of miR-195 on the proliferation, migration and angiogenesis of hEPCs under hypoxia. The results revealed that miR-195 inhibition promotes cell proliferation, migration and angiogenesis of hEPCs under hypoxia. Furthermore, GABA type A receptor associated protein like 1 (GABARAPL1) was identified as a directed target of miR-195 and GABARAPL1 silencing could decrease the effect of miR-195 knockdown on cell proliferation, migration, angiogenesis and autophagy of hEPCs under hypoxia. Together, these results indicate that miR-195 regulates cell proliferation, migration, angiogenesis and autophagy of hEPCs by targeting GABARAPL1.

Highlights

Blood clots that form within a vein are referred to as venous thrombi
The results of quantitative real-time PCR analysis (qRT-PCR) showed that the expression of miR-582, miR-195 and miR-532 was all significantly increased under conditions of hypoxia compared with that under normoxia (Figure 1)
GABARAPL1 is a direct target of miR-195 To elucidate the underlying mechanism by which miR-195 regulates cell proliferation, migration and angiogenesis of human Endothelial progenitor cells (EPCs) (hEPC) through autophagy, we explored miR-195 targets using the microRNA.org bioinformatics algorithm

Summary

Introduction

Blood clots that form within a vein are referred to as venous thrombi. The formation of a blot clot in the deep leg veins is termed deep vein thrombosis (DVT). DVT could lead to pulmonary hypertension, recurrent thrombosis or even fetal pulmonary embolism [1], and is a common complication of surgery. Many treatment options for DVT including anticoagulation, pharmacologic thrombolysis, endovascular and surgical interventions and physical measures have shown promise, they have a number of side effects, such as the risk of bleeding and wound complications [2,3]. Successful resolution of DVTrelated thrombi is important in the treatment of DVT. The development of a promising novel therapy for DVT-related thrombus resolution is essential

Methods

Results

Conclusion