TGF-β Induces Sustained Upregulation of SNAI1 and SNAI2 through Smad and Non-Smad Pathways in a Human Corneal Epithelial Cell Line
TGF-β Induces Sustained Upregulation of SNAI1 and SNAI2 through Smad and Non-Smad Pathways in a Human Corneal Epithelial Cell Line
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DOI：
10.1167/iovs.10-5635
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发表时间：
2011-04-01
影响因子：
4.4
通讯作者：
Nishio, Kazuto
中科院分区：
文献类型：
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作者：
Aomatsu, Keiichi;Arao, Tokuzo;Nishio, Kazuto
目的.本研究旨在探讨TGF-β信号诱导人角膜上皮细胞系（HCECs）上皮间质转化（EMT）相关分子表达的变化。方法.通过免疫印迹、定量实时RT-PCR和免疫荧光显微镜在HCEC中评价对TGF-β的细胞应答。结果TGF-β通过Smad和非Smad途径显著增加HCEC中SNAI 1、SNAI 2、Vim和FN 1的mRNA表达，但不增加TWIST 1的mRNA表达。间充质标志物N-钙粘蛋白的蛋白表达呈剂量依赖性增加，而上皮标志物E-钙粘蛋白的蛋白表达则被TGF-β降低。TGF-β，而不是EGF，介导的EMT样形态学变化。TGF-β和EGF都能够在短的响应时间内上调SNAI 1和SNAI 2约两倍。然而，详细的时程分析显示了截然不同的表达模式，TGF-β介导SNAI 1和SNAI 2的持续上调至少持续6天，EGF允许在8小时（类似于12小时）后恢复到基线表达值。这些数据表明，TGF-β，而不是EGF，诱导HCEC中SNAI 1和SNAI 2的持续上调。结论. TGF-β通过Smad和非Smad途径诱导HCEC中SNAI 1和SNAI 2的持续上调、EMT样形态学变化、E-钙粘蛋白的下调和N-钙粘蛋白的上调。作者的研究结果提供了深入了解TGF-β信号传导和HCEC中EMT诱导转录因子的时间表达模式。（Invest Ophthalmol维斯科学。2011; 52：2437-2443）DOI：10.1167/iovs.10-5635
PURPOSE. The aim of this study was to investigate the expression changes of epithelial mesenchymal transition (EMT)-related molecules induced by TGF-beta signaling in a human corneal epithelial cell line (HCECs). METHODS. The cellular response to TGF-beta was evaluated by immunoblotting, quantitative real-time RT-PCR, and immunofluorescence microscopy in HCECs. RESULTS. TGF-beta significantly increased mRNA expression of SNAI1, SNAI2, VIM, and FN1, but not TWIST1 through Smad and non-Smad pathways in HCECs. Protein expression of a mesenchymal marker N-cadherin was dose-dependently increased and that of an epithelial marker of E-cadherin was decreased by TGF-beta. TGF-beta, but not EGF, mediated the EMT-like morphologic changes. Both TGF-beta and EGF were capable of upregulating SNAI1 and SNAI2 by about two-fold within a short response time. However, a detailed time course analysis revealed drastically different expression patterns, with TGF-beta mediating a sustained upregulation of SNAI1 and SNAI2 for at least for 6 days and EGF allowing a return to the baseline expression values after 8 similar to 12 h. These data indicate that TGF-beta, but not EGF, induces sustained upregulation of SNAI1 and SNAI2 in HCECs. CONCLUSIONS. TGF-beta induces sustained upregulation of SNAI1 and SNAI2 through Smad and non-Smad pathways, EMT-like morphologic changes, downregulation of E-cadherin, and upregulation of N-cadherin in HCECs. The authors' findings provide insight into the TGF-beta signaling and the temporal expression patterns of EMT-inducible transcription factors in HCECs. (Invest Ophthalmol Vis Sci. 2011; 52: 2437-2443) DOI: 10.1167/iovs.10-5635