title: siRNA knockdown of tissue inhibitor of metalloproteinase-1 in keloid fibroblasts leads to degradation of collagen type I.
abstract: Keloids are defined as overgrowths of scar tissue resulting from abnormal wound healing. They are characterized by excessive dermal deposition of thick, hyalinized collagen bundles resulting from an imbalance between the production and degradation of extracellular matrix (ECM) components. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are two important regulators of ECM degradation and remodeling. To evaluate the role played by knockdown of TIMPs in keloid formation, we transduced human keloid-derived fibroblasts (KFs) with small interfering RNAs targeting TIMP-1 or -2 (siTIMP-1 or siTIMP-2) using a lentiviral vector and assessed the biological effects. We found that MMP-1/TIMP-1 and MMP-1/TIMP-2 complexes were suppressed and that MMP-2 activity was upregulated in KFs expressing siTIMP-1 or siTIMP-2. In addition, increased degradation of collagen type I was observed in the supernatant of KFs expressing siTIMP-1, but not siTIMP-2, with the suppression of cell viability and induction of apoptosis. These results suggest that targeting TIMP-1 using small interfering RNA has significant therapeutic potential as an approach to treating keloids through degradation of their thick collagen bundles.
authors.affiliation: Division of Gene Therapy Research Center for Advanced Medical Technology, Department of Biochemistry and Molecular Biology, Tokyo, Japan; Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan.
authors.forename: Masayo
authors.initials: M
authors.lastname: Aoki
authors.affiliation: Division of Gene Therapy Research Center for Advanced Medical Technology, Department of Biochemistry and Molecular Biology, Tokyo, Japan. Electronic address: kmiyake@nms.ac.jp.
authors.forename: Koichi
authors.initials: K
authors.lastname: Miyake
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan.
authors.forename: Rei
authors.initials: R
authors.lastname: Ogawa
authors.affiliation: Division of Gene Therapy Research Center for Advanced Medical Technology, Department of Biochemistry and Molecular Biology, Tokyo, Japan; Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Dohi
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Akaishi
authors.affiliation: Department of Plastic, Reconstructive and Aesthetic Surgery, Nippon Medical School, Tokyo, Japan.
authors.forename: Hiko
authors.initials: H
authors.lastname: Hyakusoku
authors.affiliation: Division of Gene Therapy Research Center for Advanced Medical Technology, Department of Biochemistry and Molecular Biology, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
mesh_terms: D017209:Apoptosis / Q000502:physiology; D024042:Collagen Type I / Q000378:metabolism*; D000090602:Collagen Type I, alpha 1 Chain; D024061:Collagen Type III / Q000378:metabolism; D017364:Collagenases / Q000378:metabolism; D003470:Culture Media / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D018093:Gelatinases / Q000378:metabolism; D006801:Humans; D007627:Keloid / Q000235:genetics / Q000473:pathology* / Q000503:physiopathology*; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D061251:Primary Cell Culture; D034741:RNA, Small Interfering / Q000235:genetics; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics* / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: C439971:COL3A1 protein, human; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D024061:Collagen Type III; D003470:Culture Media; D034741:RNA, Small Interfering; C560822:TIMP1 protein, human; C560823:TIMP2 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; D017364:Collagenases; D018093:Gelatinases
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