title: Lipopolysaccharide-induced microRNA-146a targets CARD10 and regulates angiogenesis in human umbilical vein endothelial cells.
abstract: This aim of this study was to explore the role of miRNA-146a (miR-146a) and its target genes in endothelial cells. We demonstrated that lipopolysaccharide (LPS) induced the upregulation of miR-146a in human umbilical vein endothelial cells (HUVECs), and that the induction was blocked by silencing toll-like receptors, the adaptor molecule MyD88, and the nonspecific NF-κB inhibitor BAY 11-7082. In addition, knockdown of miR-146a by transfection of the locked nucleic acid antimiR-146a significantly inhibited LPS-induced cell migration and tube formation. A combined analysis of bioinformatics miRanda algorithms and a whole genome expression microarray of immunoprecipitated Ago2 ribonucleoprotein complexes identified 14 potential target genes. Subsequent transfection with the miR-146a precursor pre-miR-146a into HUVECs validated that CARD10 was the target gene of the miR-146a, both at the mRNA and protein levels. Silencing CARD10 inhibited p65 nuclear translocation in the cells receiving LPS stimulation and increased angiogenesis. Therefore, miR-146a may play a role in regulating the angiogenesis in HUVECs by downregulating CARD10, which acts in a negative feedback regulation loop to inhibit the activation of NF-κB that normally impairs angiogenesis.
authors.affiliation: Department of Neurosurgery.
authors.forename: Cheng-Shyuan
authors.initials: CS
authors.lastname: Rau
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan.
authors.forename: Johnson Chia-Shen
authors.initials: JC
authors.lastname: Yang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan.
authors.forename: Yi-Chun
authors.initials: YC
authors.lastname: Chen
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan.
authors.forename: Chia-Jung
authors.initials: CJ
authors.lastname: Wu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan.
authors.forename: Tsu-Hsiang
authors.initials: TH
authors.lastname: Lu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan.
authors.forename: Siou-Ling
authors.initials: SL
authors.lastname: Tzeng
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan.
authors.forename: Yi-Chan
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taiwan m93chinghua@gmail.com.
authors.forename: Ching-Hua
authors.initials: CH
authors.lastname: Hsieh
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D053468:CARD Signaling Adaptor Proteins / Q000235:genetics*; D017931:DNA Primers; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D035683:MicroRNAs / Q000235:genetics*; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D016133:Polymerase Chain Reaction
chemical_list: D053468:CARD Signaling Adaptor Proteins; C430880:CARD10 protein, human; D017931:DNA Primers; D008070:Lipopolysaccharides; C542542:MIRN146 microRNA, human; C507779:MYD88 protein, human; D035683:MicroRNAs; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B
keywords: Ago2; CARD10; lipopolysaccharide (LPS); miR-146a; nuclear factor-κB (NF-κB); whole genome microarray