title: miR-483-3p controls proliferation in wounded epithelial cells.
abstract: The mechanisms that regulate keratinocyte migration and proliferation in wound healing remain largely unraveled, notably regarding possible involvements of microRNAs (miRNAs). Here we disclose up-regulation of miR-483-3p in 2 distinct models of wound healing: scratch-injured cultures of human keratinocytes and wounded skin in mice. miR-483-3p accumulation peaks at the final stage of the wound closure process, consistent with a role in the arrest of "healing" progression. Using an in vitro wound-healing model, videomicroscopy, and 5-bromo-2'-uridine incorporation, we observed that overexpression of miR-483-3p inhibits keratinocyte migration and proliferation, whereas delivery of anti-miR-483-3p oligonucleotides sustains keratinocyte proliferation beyond the closure of the wound, compared with irrelevant anti-miR treatment. Expression profiling of keratinocytes transfected with miR-483-3p identified 39 transcripts that were both predicted targets of miR-483-3p and down-regulated after miR-483-3p overexpression. Luciferase reporter assays, Western blot analyses, and silencing by specific siRNAs finally established that kinase MK2, cell proliferation marker MKI67, and transcription factor YAP1 are direct targets of miR-483-3p that control keratinocyte proliferation. miR-483-3p-mediated down-regulation of MK2, MKI67, and YAP1 thus represents a novel mechanism controlling keratinocyte growth arrest at the final steps of reepithelialization.
authors.affiliation: INSERM U634, IFR50, Faculté de Médecine, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bertero
authors.forename: Cécile
authors.initials: C
authors.lastname: Gastaldi
authors.forename: Isabelle
authors.initials: I
authors.lastname: Bourget-Ponzio
authors.forename: Véronique
authors.initials: V
authors.lastname: Imbert
authors.forename: Agnès
authors.initials: A
authors.lastname: Loubat
authors.forename: Eric
authors.initials: E
authors.lastname: Selva
authors.forename: Roser
authors.initials: R
authors.lastname: Busca
authors.forename: Bernard
authors.initials: B
authors.lastname: Mari
authors.forename: Paul
authors.initials: P
authors.lastname: Hofman
authors.forename: Pascal
authors.initials: P
authors.lastname: Barbry
authors.forename: Guerrino
authors.initials: G
authors.lastname: Meneguzzi
authors.forename: Gilles
authors.initials: G
authors.lastname: Ponzio
authors.forename: Roger
authors.initials: R
authors.lastname: Rezzonico
mesh_terms: D000818:Animals; D000906:Antibodies; D049109:Cell Proliferation; D004847:Epithelial Cells; D020868:Gene Silencing; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009841:Oligonucleotides; D012867:Skin / Q000378:metabolism; D013997:Time Factors; D014947:Wounds and Injuries / Q000378:metabolism*
chemical_list: D000906:Antibodies; C548912:MIRN483 microRNA, human; D035683:MicroRNAs; C548913:Mirn483 microRNA, mouse; D009841:Oligonucleotides; C477371:locked nucleic acid