title: Induction of keratinocyte migration via transactivation of the epidermal growth factor receptor by the antimicrobial peptide LL-37.
abstract: The closure of skin wounds is essential for resistance against microbial pathogens, and keratinocyte migration is an important step in skin wound healing. Cathelicidin hCAP18/LL-37 is an innate antimicrobial peptide that is expressed in the skin and acts to eliminate microbial pathogens. Because hCAP18/LL-37 is up-regulated at skin wound sites, we hypothesized that LL-37 induces keratinocyte migration. In this study, we found that 1 microg/ml LL-37 induced the maximum level of keratinocyte migration in the Boyden chamber assay. In addition, LL-37 phosphorylated the epidermal growth factor receptor (EGFR) after 10 min, which suggests that LL-37-induced keratinocyte migration occurs via EGFR transactivation. To test this assumption, we used inhibitors that block the sequential steps of EGFR transactivation, such as OSU8-1, CRM197, anti-EGFR no. 225 Ab, and AG1478. All of these inhibitors completely blocked LL-37-induced keratinocyte migration, which indicates that migration occurs via HB-EGF-mediated EGFR transactivation. Furthermore, CRM197, anti-EGFR no. 225, and AG1478 blocked the LL-37-induced phosphorylation of STAT3, and transfection with a dominant-negative mutant of STAT3 abolished LL-37-induced keratinocyte migration, indicating the involvement of the STAT3 pathway downstream of EGFR transactivation. Finally, we tested whether the suppressor of cytokine signaling (SOCS)/cytokine-inducible Src homology 2-containing protein (CIS) family of negative regulators of STAT3 regulates LL-37-induced keratinocyte migration. Transfection with SOCS1/Jak2 binding protein or SOCS3/CIS3 almost completely abolished LL-37-induced keratinocyte migration. In conclusion, LL-37 induces keratinocyte migration via heparin-binding-EGF-mediated transactivation of EGFR, and SOCS1/Jak 2 binding and SOCS3/CIS3 negatively regulate this migration. The results of this study suggest that LL-37 closes skin wounds by the induction of keratinocyte migration.
authors.affiliation: Department of Dermatology, Ehime University School of Medicine, Ehime, Japan.
authors.forename: Sho
authors.initials: S
authors.lastname: Tokumaru
authors.forename: Koji
authors.initials: K
authors.lastname: Sayama
authors.forename: Yuji
authors.initials: Y
authors.lastname: Shirakata
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Komatsuzawa
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Ouhara
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Hanakawa
authors.forename: Yoko
authors.initials: Y
authors.lastname: Yahata
authors.forename: Xiuju
authors.initials: X
authors.lastname: Dai
authors.forename: Mikiko
authors.initials: M
authors.lastname: Tohyama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nagai
authors.forename: Lujun
authors.initials: L
authors.lastname: Yang
authors.forename: Shigeki
authors.initials: S
authors.lastname: Higashiyama
authors.forename: Akihiko
authors.initials: A
authors.lastname: Yoshimura
authors.forename: Motoyuki
authors.initials: M
authors.lastname: Sugai
authors.forename: Koji
authors.initials: K
authors.lastname: Hashimoto
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000378:metabolism* / Q000502:physiology; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000502:physiology; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D010766:Phosphorylation; D012097:Repressor Proteins / Q000502:physiology; D000071222:Suppressor of Cytokine Signaling 1 Protein; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000502:physiology; D015533:Transcriptional Activation / Q000502:physiology*; D054804:Cathelicidins
chemical_list: D023181:Antimicrobial Cationic Peptides; D047908:Intracellular Signaling Peptides and Proteins; D012097:Repressor Proteins; C108944:SOCS1 protein, human; C106644:SOCS3 protein, human; D000071222:Suppressor of Cytokine Signaling 1 Protein; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D054804:Cathelicidins