4.6 Article

Crosstalk between Shh and TGF-β Signaling in Cyclosporine-Enhanced Cell Proliferation in Human Gingival Fibroblasts

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PLOS ONE
卷 8, 期 7, 页码 -

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PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0070128

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  1. Tri-Service General Hospital [TSGH-C98-29]
  2. National Science Council [NSC-100-2314-B-016-022-MY3]
  3. C.Y. Foundation for Advancement of Education, Sciences, and Medicine, Republic of China

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Background: Immunosuppressant cyclosporine-A induces gingival hyperplasia, which is characterized by increased fibroblast proliferation and overproduction of extracellular matrix components and regulated by transforming growth factor-beta (TGF-beta). The TGF-beta and Sonic hedgehog (Shh) pathways both mediate cell proliferation. Crosstalk between these pathways in cancer has recently been proposed, but the hierarchical pattern of this crosstalk remains unclear. In normal fibroblasts, a TGF-beta-stimulating Shh pattern was observed in induced fibrosis. However, Shh pathway involvement in cyclosporine-enhanced gingival proliferation and the existence of crosstalk with the TGF-beta pathway remain unclear. Methodology/Principal Findings: Cyclosporine enhanced mRNA and protein levels of TGF-beta and Shh in human gingival fibroblasts (RT-PCR and western blotting). A TGF-beta pathway inhibitor mitigated cyclosporine-enhanced cell proliferation and an Shh pathway inhibitor attenuated cyclosporine-enhanced proliferation in fibroblasts (MTS assay and/or RT-PCR of PCNA). Exogenous TGF-beta increased Shh expression; however, exogenous Shh did not alter TGF-beta expression. The TGF-beta pathway inhibitor mitigated cyclosporine-upregulated Shh expression, but the Shh pathway inhibitor did not alter cyclosporineupregulated TGF-beta expression. Conclusions/Significance: The TGF-beta and Shh pathways mediate cyclosporine-enhanced gingival fibroblast proliferation. Exogenous TGF-beta increased Shh expression, and inhibition of TGF-beta signaling abrogated the cyclosporine-induced upregulation of Shh expression; however, TGF-beta expression appeared unchanged by enhanced or inhibited Shh signaling. This is the first study demonstrating the role of Shh in cyclosporine-enhanced gingival cell proliferation; moreover, it defines a hierarchical crosstalk pattern in which TGF-beta regulates Shh in gingival fibroblasts. Understanding the regulation of cyclosporine-related Shh and TGF-beta signaling and crosstalk in gingival overgrowth will clarify the mechanism of cyclosporine-induced gingival enlargement and help develop targeted therapeutics for blocking these pathways, which can be applied in pre-clinical and clinical settings.

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