Journal
TISSUE & CELL
Volume 67, Issue -, Pages -Publisher
CHURCHILL LIVINGSTONE
DOI: 10.1016/j.tice.2020.101453
Keywords
Deep vein thrombosis; Endothelial progenitor cell; miR-143-3p; Autophagy-related 2B
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Deep vein thrombosis (DVT) is a common disease in vascular surgery. In recent study, microRNA (miRNA) plays a regulatory role in function of Endothelial progenitor cells (EPCs), which showed promising therapeutic choice for DVT. However, the function of miR-143-3p in EPCs remains incomplete. Flow cytometry was used to identify EPCs surface markers. Cell viability, migration, invasion and tube formation of EPCs were detected by 3-[4,5-dimethylthylthiazol-2-yl]-2,5 diphenyltetrazolium broide (MTT), wound healing, transwell and tube formation assay, respectively. TargetScan was used to predict miR-143-3p targeting genes. Dual-luciferase report assay was used to verify the interactions between miR-143-3p and autophagy-related 2B (ATG2B). Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to examine the mRNA expression levels of ATG2B and miR-143-3p. Western blot was used to examine the protein expression levels of ATG2B, LC3 and p62. The cultured EPCs showed cobblestone morphology and were identified by cell surface markers. Overexpression of miR-143-3p enhanced the viability, migration, invasion and tube formation of EPCs, but low expression of miR143-3p obtained the reverse results. ATG2B directly bound to miR-143-3p. Overexpression of miR-143-3p reduced the expression of ATG2B, but low expression of miR-143-3p increased. Overexpression of miR-143-3p decreased the expression of LC3I/II, but increased the expression of p62. Overexpression of ATG2B reversed the above-mentioned effects of EPCs which regulated by overexpression of miR-143-3p. MiR-143-3p targets ATG2B to modulate the function of EPCs and recanalization and resolution of DVT.
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