title: Formation of GW/P bodies as marker for microRNA-mediated regulation of innate immune signaling in THP-1 cells.
abstract: GW bodies (GWB or P bodies) are cytoplasmic foci thought to result from microRNA (miRNA) regulation of messenger RNA (mRNA) targets and subsequent mRNA degradation. The purpose of this study is to examine the effects of lipopolysaccharide (LPS) stimulation of human monocytes on GWB formation, miRNA induction, miRNA target regulation and downstream cytokine and chemokine expression. In response to LPS stimulation, the number of GWB consistently increased by twofold at 8 h after stimulation and this increase was abolished when the miRNA-effector proteins Rck/p54 or argonaute 2 were depleted. As the level of miR-146a increased from 19-fold up to 100-fold during LPS stimulation, the transfection of a miR-146a mimic into THP-1 cells was examined to determine whether miR-146a alone can induce similar changes in GWB. The results showed transfected miR-146a could produce a comparable increase in the number of GWB and this was accompanied by a reduction in major cytokines/chemokines induced by LPS. These data show that the increase in size and number of GWB may serve as a biomarker for miRNA-mediated gene regulation, and miR-146a has a significant role in the regulation of LPS-induced cytokine production in THP-1 cells.
authors.affiliation: Department of Oral Biology, University of Florida, Gainesville, FL 32610-0424, USA.
authors.forename: Kaleb M
authors.initials: KM
authors.lastname: Pauley
authors.forename: Minoru
authors.initials: M
authors.lastname: Satoh
authors.forename: Brad A
authors.initials: BA
authors.lastname: Pauley
authors.forename: Paul R
authors.initials: PR
authors.lastname: Dominguez-Gutierrez
authors.forename: Shannon M
authors.initials: SM
authors.lastname: Wallet
authors.forename: L Shannon
authors.initials: LS
authors.lastname: Holliday
authors.forename: Seunghee
authors.initials: S
authors.lastname: Cha
authors.forename: Westley H
authors.initials: WH
authors.lastname: Reeves
authors.forename: Edward K L
authors.initials: EK
authors.lastname: Chan
mesh_terms: D060565:Argonaute Proteins; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D018925:Chemokines / Q000096:biosynthesis; D022083:Cytoplasmic Structures / Q000187:drug effects / Q000276:immunology*; D053487:DEAD-box RNA Helicases / Q000172:deficiency / Q000378:metabolism; D015852:Eukaryotic Initiation Factor-2 / Q000172:deficiency / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects / Q000276:immunology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D018448:Models, Immunological; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D011518:Proto-Oncogene Proteins / Q000172:deficiency / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology*; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D015415:Biomarkers; D018925:Chemokines; D015852:Eukaryotic Initiation Factor-2; D008070:Lipopolysaccharides; D035683:MicroRNAs; D011518:Proto-Oncogene Proteins; C094683:DDX6 protein, human; D053487:DEAD-box RNA Helicases