title: Rho plays a key role in TGF-β1-induced proliferation and cytoskeleton rearrangement of human periodontal ligament cells.
abstract: Human periodontal ligament cells (hPDLCs) form specialised connective tissues that influence the lifespan of the tooth. Periodontal disease is a chronic infectious disease of the periodontal supporting tissues caused by a variety of factors, particularly the loss of hPDLCs. Transforming growth factor-β1 (TGF-β1) is a multifunctional cytokine known to play an important role in periodontal disease, but little is known about the effects of TGF-β1 on human PDL cells. To determine how TGF-β1 mediates the changes in hPDLCs, we characterised the effects of TGF-β1 treatment on hPDLCs. We then elucidated the signalling pathway that mediates these effects. Serum-starved hPDLCs were incubated with 10ng/mL TGF-β1, and their proliferation was examined using the Cell Counting Kit-8, while their morphological changes were examined by phase-contrast microscopy. F-actin reorganisation was visualised by phalloidin staining and confocal microscopy. Protein expression was analysed by western blotting. We found that TGF-β1 treatment induced proliferation and cytoskeletal reorganisation, decreased Rho-GDIa protein expression, activated ROCK protein expression, and increased the phosphorylation of LIM kinase and cofilin. Proliferation and cytoskeletal rearrangement were suppressed by pre-treatment with the ROCK inhibitor Y-27632; additionally, expression of ROCK protein and phosphorylation of LIM kinase and cofilin were decreased by Y-27632, while Rho-GDIa knockdown by targeted siRNA transfection causes opposite effects. Therefore, we propose that TGF-β1 induces proliferation and cytoskeletal rearrangement in hPDLCs via Rho GTPase-dependent pathways that modulate ROCK, LIM kinase, and cofilin activity.
authors.affiliation: Department of Stomatology, Shanghai First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Stomatology, Central Hospital of Minhang District, Shanghai, China.
authors.forename: Tingle
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Stomatology, Shanghai First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. Electronic address: smgjzh160@sohu.com.
authors.forename: Meng
authors.initials: M
authors.lastname: Song
authors.affiliation: Department of Stomatology, Shanghai First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. Electronic address: pip0003@163.com.
authors.forename: Jinsong
authors.initials: J
authors.lastname: Pan
mesh_terms: D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects*; D006801:Humans; D010513:Periodontal Ligament / Q000166:cytology*; D010641:Phenotype; D015398:Signal Transduction / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D020742:rhoA GTP-Binding Protein / Q000502:physiology*
chemical_list: D053773:Transforming Growth Factor beta1; D020742:rhoA GTP-Binding Protein
keywords: Cytoskeletal rearrangement; Human periodontal ligament cells; Proliferation; Rho signalling pathway; TGF-β1