The role of Smad3 in the fibrotic phenotype in human vocal fold fibroblasts.
The role of Smad3 in the fibrotic phenotype in human vocal fold fibroblasts.
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DOI：
10.1002/lary.25673
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发表时间：
2016-05
期刊：
影响因子：
--
通讯作者：
Amin MR
中科院分区：
文献类型：
--
作者：
Branski RC;Bing R;Kraja I;Amin MR
目的 研究 Smad3 作为转化生长因子 (TGF)-β1 介导的与正常人声带成纤维细胞纤维化相关的细胞活性调节剂的作用。我们还试图通过 siRNA 确认 Smad3 敲低的时间稳定性。采用声带成纤维细胞来确定 Smad3 敲低对 TGF-β1 介导的迁移和收缩以及结缔组织生长因子 (CTGF) 调节的影响。基于 Smad3 在纤维化中的作用，我们假设 Smad3 是体内治疗操作的理想候选者。在我们的正常人声带细胞系中通过 siRNA 体外敲除 Smad3。采用三维胶原凝胶收缩和划痕测定分别确定 Smad3 对 TGF-β1 介导的收缩和迁移的作用。 Smad3 在 CTGF 诱导中的作用通过 SDS-Page 进行了表征。 Smad3 信号传导对 Smad7 mRNA 和蛋白质的影响也进行了量化。 Smad3 敲除在长达 72 小时内保持暂时稳定 (p<0.001)。 Smad3 敲除减少了 TGF-β1 介导的胶原凝胶收缩和迁移。 Smad3 敲低也会减弱 CTGF 的诱导，但对 TGF-β1 介导的 Smad7 mRNA 或蛋白诱导没有影响。 TGF-β1 刺激我们细胞系中的促纤维化细胞活性，并且这些活性随着 Smad3 敲低而大大降低。这些数据为 Smad3 治疗声带纤维化的靶向治疗提供了持续的支持，因为它似乎可以调节纤维化表型。
To investigate the role of Smad3 as a regulator of transforming growth factor (TGF)-β1-mediated cell activities associated with fibrosis in normal human vocal fold fibroblasts. We also sought to confirm the temporal stability of Smad3 knockdown via siRNA. Vocal fold fibroblasts were employed to determine the effects of Smad3 knockdown on TGF-β1-mediated migration and contraction as well as regulation of connective tissue growth factor (CTGF). We hypothesized that Smad3 is an ideal candidate for therapeutic manipulation in vivo based on its role in fibrosis. In vitro Knockdown of Smad3 via siRNA was performed in our normal human vocal fold cell line. Three-dimensional collagen gel contraction and scratch assays were employed to determine the role of Smad3 on TGF-β1-mediated contraction and migration, respectively. The role Smad3 in the induction of CTGF was characterized via SDS-Page. The effects of Smad3 signaling on Smad7 mRNA and protein were also quantified. Smad3 knockdown was temporally stable up to 72 hours (p<0.001). Smad3 knockdown diminished TGF-β1-mediated collagen gel contraction and migration. Smad3 knockdown also blunted induction of CTGF, but had no effect on TGF-β1-mediated Smad7 mRNA or protein induction. TGF-β1 stimulated pro-fibrotic cell activities in our cell line and these actions were largely reduced with Smad3 knockdown. These data provide continued support for therapeutic targeting of Smad3 for vocal fold fibrosis as it appears to regulate the fibrotic phenotype.
影响因子：
2.6
作者：
Sandulache, Vlad C.;Singh, Tripti;Li-Korotky, Ha Sheng;Lo, Chia Y.;Otteson, Todd D.;Barsic, Mark;Dohar, Joseph E.;Hebda, Patricia A.
通讯作者：
Hebda, Patricia A.