TY - JOUR T1 - Knockdown of GAS5 restores ox-LDL-induced impaired autophagy flux via upregulating miR-26a in human endothelial cells JF - bioRxiv DO - 10.1101/293324 SP - 293324 AU - Weijie Liang AU - Taibing Fan AU - Lin Liu AU - Lianzhong Zhang Y1 - 2018/01/01 UR - http://biorxiv.org/content/early/2018/04/02/293324.abstract N2 - Background Oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell (EC) injury and autophagy dysfunction play a vital role in the development of atherosclerosis. LncRNAs have been identified to participate in the regulation of pathogenesis of atherosclerosis. However, it remains largely undefined whether growth-arrest specific transcript 5 (GAS5) could influence ox-LDL-induced autophagy dysfunction in ECs.Methods The expressions of GAS5 and miR-26a in the plasma samples of patients with atherosclerosis and ox-LDL-treated human aortic endothelial cells (HAECs) were detected by qRT-PCR. Luciferase reporter assay, RNA immunoprecipitation (RIP), and RNA pull down were performed to validate whether GAS5 could directly interact with miR-26a. The effects of ox-LDL, GAS5 or combined with miR-26a on apoptosis and autophagy were evaluated by flow cytometry analysis and western blot, respectively.Results GAS5 expression was upregulated and miR-26a was downregulated in the plasma samples of patients with atherosclerosis and ox-LDL-treated HAECs. There was reciprocal inhibition between GAS5 and miR-26a expressions in ox-LDL-treated HAECs. We further demonstrated that GAS5 directly interacted with miR-26a in ox-LDL-treated HAECs. Additionally, ox-LDL administration induced apoptosis and impaired autophagy flux in HAECs. Rescue experiments demonstrated that GAS5 knockdown restored ox-LDL-induced impaired autophagy flux by upregulating miR-26a in HAECs.Conclusion Knockdown of GAS5 restores ox-LDL-induced impaired autophagy flux via upregulating miR-26a in human endothelial cells, revealing a novel regulatory mechanism for ox-LDL-induced impaired autophagy flux in ECs through ceRNA crosstalk.ox-LDLOxidized low-density lipoproteinECendothelial cellGAS5growth-arrest specific transcript 5HAECshuman aortic endothelial cellsRIPRNA immunoprecipitationncRNAsnoncoding RNAslncRNAlong noncoding RNAmiRNAmicroRNAHFDhigh-fat dietapoEapolipoprotein EceRNAcompetitive endogenousRNA miR-NCmiRNA negative controlcDNAcomplementary DNAsnRNAsmall nuclear RNASDS-PAGEsodium dodecyl sulfate-polyacrylamide gelTBSTTris-based saline with Tween 20RISCRNA-induced silencing complexTSLPthymic stromal lymphopoietinHA-VSMCshuman aorta vascular smooth muscle cellsCADcoronary artery disease ER -