title: High mobility group box 1 induced human lung myofibroblasts differentiation and enhanced migration by activation of MMP-9.
abstract: High mobility group box 1 (HMGB1) is a nuclear protein that involves the binding with DNA and influences chromatin regulation and transcription. HMGB1 is also a cytokine that can activate monocytes and neutrophils involved in inflammation. In this study, we investigated the role of HMGB1 on cellular activation using human fibroblast cell line WI-38. After treatment with 1, 10, and 100 ng/mL of HMGB1 for 24 h, we did not find obviously cytotoxicity and cellular proliferation of WI-38 cells by MTT and BrdU incorporation assay, respectively. However, we found that treatment with 10 and 100 ng/mL of HMGB1 induced the differentiation of lung fibroblasts into myofibroblasts and myofibroblasts showed higher migration ability through activation of matrix metalloproteinase (MMP)-9 activation. To delineate the mechanism underlying HMGB1-induced cellular migration, we examined HMGB1-induced mitogen activated protein kinases (MAPKs), including extracellular signal related kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen activated protein kinase (p38) phosphorylation, as well as nuclear factor (NF)-κB nuclear translocation. Using specific inhibitors and shRNAs of protein kinases, we observed that repression of ERK, JNK, p38, and NF-κB all inhibited HMGB1-induced cellular differentiation, migration and MMP-9 activation in WI-38 cells. In addition, knocking down of RAGE but not TLR2 and TLR4 by shRNAs attenuated HMGB1-induced myofibroblast differentiation and migration. In conclusion, our study demonstrated that HMGB1 induced lung fibroblasts' differentiation into myofibroblasts and enhanced cell migration through induction of MMP-9 activation and the RAGE-MAPK and NF-κB interaction signaling pathways. Targeting HMGB1 might be a potential therapeutic approach for alleviation of airway remodeling seen in chronic airway inflammatory diseases.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, China Medical University, Taichung, Taiwan; Graduate Institute of Basic Medical Science, College of Medicine, China Medical University, Taichung, Taiwan.
authors.forename: Chen-Chen
authors.initials: CC
authors.lastname: Lee
authors.affiliation: Graduate Institute of Basic Medical Science, College of Medicine, China Medical University, Taichung, Taiwan.
authors.forename: Chien-Neng
authors.initials: CN
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
authors.forename: Yueh-Lun
authors.initials: YL
authors.lastname: Lee
authors.affiliation: Graduate Institute of Basic Medical Science, College of Medicine, China Medical University, Taichung, Taiwan.
authors.forename: Yi-Ru
authors.initials: YR
authors.lastname: Tsai
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, China Medical University, Taichung, Taiwan.
authors.forename: Jau-Jin
authors.initials: JJ
authors.lastname: Liu
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology; D024243:HMGB1 Protein / Q000494:pharmacology*; D006801:Humans; D008168:Lung / Q000166:cytology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D058628:Myofibroblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000378:metabolism
chemical_list: D024243:HMGB1 Protein; D016328:NF-kappa B; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D020780:Matrix Metalloproteinase 9