title: Cyclosporine A promotes cell proliferation, collagen and α-smooth muscle actin expressions in rat gingival fibroblasts by Smad3 activation and miR-29b suppression.
abstract: OBJECTIVE
  Transforming growth factor (TGF)-β/Smad3 signaling promotes tissue fibrosis, and miR-29b is a downstream inhibitor of TGF-β/Smad3-mediated fibrosis, both of which may be involved in the pathogenesis of cyclosporine A-induced gingival overgrowth. The aim of this study is to investigate the effect of miR-29b and TGF-β/Smad3 signaling in cyclosporine A-stimulated rat gingival fibroblasts.
  
  
  METHODS
  We explored the effect of cyclosporine A on cell proliferation, type Ι collagen (COL1) and alpha-smooth muscle actin (α-SMA) expressions in rat gingival fibroblasts with the Cell Counting Kit-8 assay, real-time polymerase chain reaction, western blot and immunofluorescence. The expressions of TGF-β1, Smad3 and miR-29b were also evaluated after cyclosporine A treatment. The effects of Smad3 and miR-29b on cyclosporine A-induced alterations were further determined by Smad3 knockdown and miR-29b overexpression using transient transfection experiments.
  
  
  RESULTS
  We found that cyclosporine A increased cell proliferation, COL1 and α-SMA expressions in rat gingival fibroblasts. TGF-β1 mRNA and protein expressions were upregulated and phosphorylated Smad3 was activated by cyclosporine A treatment, while miR-29b expression was significantly suppressed. Moreover, Smad3 knockdown and miR-29b overexpression reversed cyclosporine A-enhanced cell proliferation, COL1 and α-SMA expressions in gingival fibroblasts. In addition, the suppression of miR-29b upon cyclosporine A exposure was significantly elevated by Smad3 knockdown, whereas cyclosporine A-induced Smad3 activation was not altered by miR-29b overexpression.
  
  
  CONCLUSIONS
  These results suggest that cyclosporine A promotes cell proliferation, collagen and α-SMA expressions in rat gingival fibroblasts by Smad3 activation and miR-29b suppression.
authors.affiliation: Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: F
authors.initials: F
authors.lastname: Yang
authors.affiliation: Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: X
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Gong
mesh_terms: D000199:Actins / Q000096:biosynthesis*; D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects*; D003094:Collagen / Q000096:biosynthesis*; D016572:Cyclosporine / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005881:Gingiva / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D051900:Smad3 Protein / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D000199:Actins; C526366:MIRN29 microRNA, rat; D035683:MicroRNAs; D051900:Smad3 Protein; C497787:Smad3 protein, rat; D053773:Transforming Growth Factor beta1; D016572:Cyclosporine; D003094:Collagen
keywords: Smad3; cyclosporine A; gingival overgrowth; miR-29b; transforming growth factor