title: Induction of matrix metalloproteinase-1 by small interfering RNA targeting connective tissue growth factor in dermal fibroblasts from patients with systemic sclerosis.
abstract: We aimed to evaluate the effect of small interfering RNA (siRNA) targeting CTGF on extracellular matrices (ECMs) metabolism in normal and SSc fibroblasts. Normal and SSc fibroblasts were transfected with CTGF-specific siRNAs to silence CTGF synthesis. After silencing CTGF, production of type I collagen and matrix metalloproteinase (MMP)-1 by fibroblasts stimulated with TGF-beta was examined. Then quantitative analyses of protein production or mRNA expression of type I collagen, MMP-1,-2,-9 and tissue inhibitor of metalloproteinase (TIMP)-1 with TGF-beta stimulation were carried out. Furthermore, after silencing CTGF, proliferations of normal and SSc fibroblasts were investigated. CTGF-specific siRNA significantly reduced CTGF production. The production of type I collagen was significantly reduced by CTGF silencing in normal fibroblasts. The CTGF silencing significantly increased the production of MMP-1 and decreased the production of TIMP-1 in SSc fibroblasts. The mRNA expression of MMP-1 was increased in CTGF-silenced SSc fibroblasts, but not in normal fibroblasts. There were no significant changes in the production or mRNA expression of other ECM-related genes in normal and SSc fibroblasts. Fibroblast proliferations were suppressed by CTGF silencing in normal and SSc fibroblasts. Our data showed that MMP-1 was increased by CTGF-specific siRNA transfection only in SSc fibroblasts. RNAi targeting CTGF could be a novel therapeutic strategy for SSc.
authors.affiliation: Department of Dermatology, Gunma University Graduate School of Medicine, Maebashi, Japan.
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Ishibuchi
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Abe
authors.forename: Yoko
authors.initials: Y
authors.lastname: Yokoyama
authors.forename: Osamu
authors.initials: O
authors.lastname: Ishikawa
mesh_terms: D016022:Case-Control Studies; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism*; D020405:Dermis / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D020868:Gene Silencing; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000494:pharmacology* / Q000627:therapeutic use; D012595:Scleroderma, Systemic / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: C526317:CCN2 protein, human; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta; D055513:Connective Tissue Growth Factor; D020781:Matrix Metalloproteinase 1