Abstract
Background Numerous studies have highlighted that long noncoding RNA (lncRNA) can indirectly regulate the expression of mRNAs by binding to microRNA (miRNA). LncRNA-associated ceRNA networks play a vital role in the initiation and progression of several pathological mechanisms. However, the lncRNA-miRNA-mRNA ceRNA network in endothelial cells under cyclic stretch is seldom studied. Methods The miRNA, mRNA, and lncRNA expression profiles of 6 human umbilical vein endothelial cells (HUVECs) under circumferential stress were obtained by next-generation sequencing (NGS). We identified the differential expression of miRNAs, mRNAs, and lncRNAs using the R software package GDCRNATools. Cytoscape was adopted to construct a lncRNA-miRNA-mRNA ceRNA network. In addition, through GO and KEGG pathway annotations, we analyzed gene functions and their related pathways. We also adopted ELISA and TUNEL to investigate the effect of si-NEAT1 on endothelial inflammation and apoptosis. Results We recognized a total of 32978 lncRNAs, 1046 miRNAs, and 31958 mRNAs in 6 samples; among them, 155 different expressed lncRNAs, 74 different expressed miRNAs, and 960 different mRNAs were adopted. Based on the established theory, the ceRNA network was composed of 13 lncRNAs, 44 miRNAs, and 115 mRNAs. We constructed and visualized a lncRNA-miRNA-mRNA network, and the top 20 nodes are identified after calculating their degrees. The nodes with most degrees in three kinds of RNAs are hsa-miR-4739, NEAT1, and MAP3K2. Functional analysis showed that different biological processes enriched in biological regulation, response to stimulus and cell communication. Pathway analysis was mainly enriched in longevity regulating, cell cycle, mTOR, and FoxO signaling pathway. Circumferential stress can significantly downregulate NEAT1, and after transducing si-NEAT1 for 24 h, inflammatory cytokine IL-6 and MCP-1 were significantly increased; furthermore, fewer TUNEL-positive cells were found in the si-NEAT1 treated group. Conclusions The establishing of a ceRNA network can help further understand the mechanism of vein graft failure. Our data demonstrated that NEAT1 may be a core factor among the mechanical stress factors and that cyclic stress can significantly reduce expression of NEAT1, give rise to inflammation in the early stage of endothelial dysfunction, and promote EC apoptosis, which may play an essential role in vein graft failure.
Highlights
Coronary Artery Bypass Grafting (CABG) is an economical and effective treatment for most cases of multiple or left main coronary arteries [1]
We adopted a computer-controlled circumferential stress unit (Flexcell 5100, Flexcell International Corporation, McKeesport, PA) to compose cyclic stretch to human umbilical vein endothelial cells (HUVECs) for 24 hours including a condition of cyclic deformation at 60 cycles/min and elongation at 18%. e control group was maintained in the 6-well plate under the same condition but without mechanical stretch
After processing the raw data, we identified a total of 32978 long noncoding RNA (lncRNA), 1046 miRNAs, and 31958 mRNAs in 6 samples of endothelial cells compared with the established databases
Summary
Coronary Artery Bypass Grafting (CABG) is an economical and effective treatment for most cases of multiple or left main coronary arteries [1]. After the vein was transplanted from the venous system to the arterial system, the inflammation and apoptosis of ECs could be induced by changing the mechanical environment, which plays an important role in several pathological processes, including intimal hyperplasia, atherosclerosis, and occlusion [7]. The lncRNA-miRNA-mRNA ceRNA network in endothelial cells under cyclic stretch is seldom studied. E miRNA, mRNA, and lncRNA expression profiles of 6 human umbilical vein endothelial cells (HUVECs) under circumferential stress were obtained by next-generation sequencing (NGS). Cytoscape was adopted to construct a lncRNA-miRNA-mRNA ceRNA network. Our data demonstrated that NEAT1 may be a core factor among the mechanical stress factors and that cyclic stress can significantly reduce expression of NEAT1, give rise to inflammation in the early stage of endothelial dysfunction, and promote EC apoptosis, which may play an essential role in vein graft failure
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