期刊
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
卷 54, 期 10, 页码 6456-6462出版社
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
DOI: 10.1167/iovs.13-12669
关键词
glaucoma; TGF-beta; trabecular meshwork
资金
- Interdisziplinares Zentrum fur Klinische Forschung Wurzburg (IZKF Seed Grant) [Z 4-81]
- Deutsche Forschungsgemeinschaft [DFG Schl 563/3]
PURPOSE. The effects of TGF-beta 2 on cadherin-mediated cell-cell adhesion in human trabecular meshwork (HTM) cells were characterized, since TGF-beta-induced changes in the cytoskeleton, cell-cell, and cell-matrix interactions of HTM cells are suggested to have a significant role in primary open angle glaucoma. METHODS. The HTM cells were derived from donor cornea rings and treated with TGF-beta 2 or vehicle, and protein expression was studied by Western Blot, while protein localization was studied by fractionation of lysates and by confocal immunofluorescence microscopy. Cell-cell adhesion was assessed functionally in dissociation experiments and N-cadherin-mediated cell contact formation in cell spreading experiments on cadherin-coated substrates. A rho-associated protein kinase (ROCK) inhibitor was used to evaluate the contribution of cytoskeletal tension to TGF-beta 2-induced changes in protein expression and localization. RESULTS. TGF-beta 2 activated Smad-2/3, serine-threonine kinase (AKT), and extracellular signal-regulated kinase (ERK) signaling, and enhanced expression of beta-catenin as well as N- and OB-cadherin. The nuclear fraction of beta-catenin was not enhanced by TGF-beta 2. Immunofluorescence microscopy revealed an increased localization of N-cadherin and beta-catenin to cell-cell adhesions, and an increase in F-actin. The TGF-beta 2 increased cell-cell adhesion strength and enhanced N-cadherin-mediated cell contact formation. This effect was blocked by inhibition of mitogen-activated protein kinase kinase (MEK) or AKT. Cytoskeletal relaxation by a ROCK inhibitor did not prevent a TGF-beta 2-induced increase in cadherin and beta-catenin expression. CONCLUSIONS. The cytokine TGF-beta 2 enhances cadherin-mediated cell-cell adhesion and beta-catenin expression in HTM cells. Increased cell-cell adhesion may contribute to biomechanical alterations in glaucomatous trabecular meshwork (TM), and changes in beta-catenin levels and its possible sequestration to cell adhesion sites may affect Wnt signaling. Thus, the crosstalk of TGF-beta 2 and Wnt signaling in TM may deserve further investigation.
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