Wnt signaling induces epithelial-mesenchymal transition with proliferation in ARPE-19 cells upon loss of contact inhibition.
Wnt signaling induces epithelial-mesenchymal transition with proliferation in ARPE-19 cells upon loss of contact inhibition.
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DOI：
10.1038/labinvest.2011.201
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发表时间：
2012-05
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影响因子：
--
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视网膜色素上皮（RPE）的增殖和上皮间质转化（EMT）是增殖性玻璃体视网膜病变的标志。本研究旨在阐明生长因子，即表皮生长因子（EGF）、成纤维细胞生长因子-2（FGF-2）和转化生长因子-β1（TGF-β1）在控制RPE在经历EMT时如何增殖中的作用。当融合后ARPE-19细胞的接触抑制被EGTA破坏时，仅在EGF和/或FGF-2存在的情况下注意到BrdU标记的增加，并且伴随EMT，如正常RPE表型的丧失所证明的（RPE 65、N-钙粘蛋白、ZO-1和Na，K-ATP酶的细胞定位改变）和间充质表型的获得（波形蛋白、S100 A4和α-SMA表达增加）。EGTA + EGF+FGF-2的EMT伴随着典型Wnt信号的激活（通过TCF/LEF启动子活性、β-连环蛋白和LEF 1蛋白的核水平增加和它们之间的相互作用以及通过β-连环蛋白的过表达进行的复制来判断），通过同时加入XAV 939（一种Wnt抑制剂）来消除，但与Hippo信号传导的抑制无关（核TAZ或雅普和细胞质p-TAZ或p-雅普的阴性表达）。通过EGTA处理稳定的S33 Y β-catenin的过表达证实了Wnt信号传导对EMT与增殖的致病作用。此外，在TGF-β1存在下，EGTA破坏的接触抑制也导致EMT，但抑制增殖和Wnt信号传导。EGF+FGF-2可有效激活Wnt信号通路，并与TGF-β1协同激活Smad/ZEB 1/2信号通路。这些发现建立了一个框架，进一步解剖如何RPE可能参与了一些增殖性玻璃体视网膜病变的特点是EMT。
Proliferation and epithelial-mesenchymal transition (EMT) of the retinal pigment epithelium (RPE) are hallmarks of proliferative vitreoretinopathy. This study aims at clarifying the role of growth factors, i.e. epidermal growth factor (EGF), fibroblast growth factor-2 (FGF-2), and transforming growth factor-β1 (TGF-β1) in controlling how RPE proliferates while undergoing EMT. When contact inhibition of post-confluent ARPE-19 cells was disrupted by EGTA, an increase of BrdU labeling was noted only in the presence of EGF and/or FGF-2, and was accompanied by EMT as evidenced by the loss of a normal RPE phenotype (altered cytolocalization of RPE65, N-cadherin, ZO-1, and Na,K-ATPase) and the gain of a mesenchymal phenotype (increased expression of vimentin, S100A4, and α-SMA). EMT with proliferation by EGTA plus EGF+FGF-2 was accompanied by activation of canonical Wnt signaling (judged by the TCF/LEF promoter activity, increased nuclear levels of and interaction between β-catenin and LEF1 proteins, and the replication by overexpression of β-catenin), abolished by concomitant addition of XAV939, a Wnt inhibitor, but not associated with suppression of Hippo signaling (negative expression of nuclear TAZ or YAP and cytoplasmic p-TAZ or p-YAP). The causative role of Wnt signaling on EMT with proliferation was confirmed by overexpression of stable S33Y β-catenin with EGTA treatment. In addition, contact inhibition disrupted by EGTA in the presence of TGF-β1 also led to EMT but suppressed proliferation and Wnt signaling. The Wnt signaling triggered by EGF+FGF-2 was sufficient and synergized with TGF-β1 in activating the Smad/ZEB1/2 signaling responsible for EMT. These findings establish a framework for further dissecting how RPE might partake in a number of proliferative vitreoretinopathies characterized by EMT.
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DOI：
10.1126/science.1199010
发表时间：
2011-04-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Heallen T;Zhang M;Wang J;Bonilla-Claudio M;Klysik E;Johnson RL;Martin JF
通讯作者：
Martin JF
影响因子：
4.4
作者：
Burke, Janice M.;Hong, Jeehee
通讯作者：
Hong, Jeehee
影响因子：
2.1
作者：
BAUDOUIN, C;FREDJREYGROBELLET, D;GASTAUD, P
通讯作者：
GASTAUD, P
影响因子：
4.8
作者：
Cushing, Melinda C.;Mariner, Peter D.;Liao, Jo-Tsu;Sims, Evan A.;Anseth, Kristi S.
通讯作者：
Anseth, Kristi S.
影响因子：
11.8
作者：
Ciruna, B;Rossant, J
通讯作者：
Rossant, J