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16 September 2019 : Animal Research  

Identification and Analysis of Differentially Expressed Genes in Human Saphenous Vein Endothelial Cells Overexpressing Domain-Containing mTOR-Interacting Protein (DEPTOR) by RNA-Seq

Wenjun Wang1ABCE, Yiying Li2BD, Qun Wang1BCDF, Yuanping Cao1CDE, Fudong Wang1BDEF, Wan Li1ABDEFG*

DOI: 10.12659/MSM.915442

Med Sci Monit 2019; 25:6965-6971

Abstract

BACKGROUND: Autologous saphenous vein is the most common choice for coronary artery bypass grafting. This study was conducted to identify and characterize differentially expressed genes (DEGs) induced by overexpressing DEPTOR in human saphenous vein endothelial cells (hsVECs) that might play roles in restenosis.

MATERIAL AND METHODS: hsVECs isolated from the saphenous veins were transfected with DEPTOR overexpression vector and analyzed for mTOR expression. RNA was prepared from the cells and sequenced using high-throughput sequencing technology (RNA-Seq). The DEGs were analyzed based on enrichment scores in GO terms and KEGG pathways.

RESULTS: The cells had typical hsVEC morphology and characteristics based on the HE staining and immunohistochemical and immunofluorescence assays. The expression of mTOR increased, and 102 genes were upregulated, and 409 genes were downregulated after DEPTOR overexpression. KEGG analysis showed that the DEGs were mainly enriched in 20 signal pathways, such as Focal adhesion and ECM-receptor interaction pathways. The DEGs were enriched in GO terms such as integrin binding and glycosaminoglycan binding. For cellular components, GO analysis revealed that the DEGs were enriched in main axon, plasma membrane part, cell junction, and proteinaceous extracellular matrix. DEGs included many cytokines, such as bone morphogenetic protein-7, interleukin-8, interleukin-1β, and inhibin, which have important effects on vascular growth and inflammation.

CONCLUSIONS: The overexpression of DEPTOR in hsVECs results in DEGs that are involved in cell proliferation and differentiation, intercellular junction, and extracellular matrix receptor. These findings may provide valuable molecular information for improving venous permeability through manipulation of DEPTOR and related mTOR pathways.

Keywords: Coronary Artery Bypass, Coronary Restenosis, endothelial cells, Gene Expression Profiling, Gene Expression Regulation, gene ontology, Intracellular Signaling Peptides and Proteins, RNA-Seq, Saphenous Vein, TOR Serine-Threonine Kinases

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750