title: Regulation of NF-κB signaling by oxidized glycerophospholipid and IL-1β induced miRs-21-3p and -27a-5p in human aortic endothelial cells.
abstract: Exposure of endothelial cells (ECs) to agents such as oxidized glycerophospholipids (oxGPs) and cytokines, known to accumulate in atherosclerotic lesions, perturbs the expression of hundreds of genes in ECs involved in inflammatory and other biological processes. We hypothesized that microRNAs (miRNAs) are involved in regulating the inflammatory response in human aortic endothelial cells (HAECs) in response to oxGPs and interleukin 1β (IL-1β). Using next-generation sequencing and RT-quantitative PCR, we characterized the profile of expressed miRNAs in HAECs pre- and postexposure to oxGPs. Using this data, we identified miR-21-3p and miR-27a-5p to be induced 3- to 4-fold in response to oxGP and IL-1β treatment compared with control treatment. Transient overexpression of miR-21-3p and miR-27a-5p resulted in the downregulation of 1,253 genes with 922 genes overlapping between the two miRNAs. Gene Ontology functional enrichment analysis predicted that the two miRNAs were involved in the regulation of nuclear factor κB (NF-κB) signaling. Overexpression of these two miRNAs leads to changes in p65 nuclear translocation. Using 3' untranslated region luciferase assay, we identified 20 genes within the NF-κB signaling cascade as putative targets of miRs-21-3p and -27a-5p, implicating these two miRNAs as modulators of NF-κB signaling in ECs.
authors.affiliation: Departments of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Milagros C
authors.initials: MC
authors.lastname: Romay
authors.affiliation: Medicine, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Nam
authors.initials: N
authors.lastname: Che
authors.affiliation: Departments of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Scott N
authors.initials: SN
authors.lastname: Becker
authors.affiliation: Medicine, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Delila
authors.initials: D
authors.lastname: Pouldar
authors.affiliation: Human Genetics, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Raffi
authors.initials: R
authors.lastname: Hagopian
authors.affiliation: Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Xinshu
authors.initials: X
authors.lastname: Xiao
authors.affiliation: Departments of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095 Medicine, University of California, Los Angeles, Los Angeles, CA 90095 Human Genetics, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Aldons J
authors.initials: AJ
authors.lastname: Lusis
authors.affiliation: Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Judith A
authors.initials: JA
authors.lastname: Berliner
authors.affiliation: Medicine, University of California, Los Angeles, Los Angeles, CA 90095.
authors.forename: Mete
authors.initials: M
authors.lastname: Civelek
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D035683:MicroRNAs / Q000235:genetics*; D010084:Oxidation-Reduction; D010713:Phosphatidylcholines / Q000737:chemistry / Q000494:pharmacology*; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000187:drug effects*; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: C468114:1-palmitoyl-2-arachidonyl-3-phosphorylcholine; D020413:3' Untranslated Regions; D053583:Interleukin-1beta; C524940:MIRN21 microRNA, human; C531388:MIRN27 microRNA, human; D035683:MicroRNAs; D010713:Phosphatidylcholines; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha
keywords: cell signaling; cytokines; gene expression; inflammation oxidized lipids; interleukin 1β; microRNAs; nuclear factor κB