Cytoskeletal re-arrangement in TGF-β1-induced alveolar epithelial-mesenchymal transition studied by atomic force microscopy and high-content analysis
Cytoskeletal re-arrangement in TGF-β1-induced alveolar epithelial-mesenchymal transition studied by atomic force microscopy and high-content analysis
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DOI：
10.1016/j.nano.2011.06.021
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发表时间：
2012-04-01
影响因子：
5.4
通讯作者：
Ehrhardt, Carsten
中科院分区：
文献类型：
--
作者：
Buckley, Stephen T.;Medina, Carlos;Ehrhardt, Carsten
上皮-间质转化（EMT）与特发性肺纤维化的发病机制密切相关。与这种表型转变相关的是获得细长的细胞形态和建立应力纤维。这些EMT相关变化对细胞力学的影响程度尚不清楚。我们评估了接触TGF-β 1后肺泡上皮细胞（A549）的生物力学特性。使用原子力显微镜，细胞刚度和表面膜特征的变化进行了测定。用TGF-β 1刺激引起刚度的显著增加，其通过胶原蛋白I基质增强。此外，TGF-β 1处理的细胞表现出更粗糙的表面轮廓，具有显著的突起。使用基于图像的高含量分析系统的刺激细胞的形态学属性的同时定量检查揭示了细胞形状，F-肌动蛋白含量和分布的显着变化。总之，这些研究指出了细胞间质相关的细胞结构和进行EMT的肺泡上皮细胞的机械动力学之间的强相关性。
Epithelial-mesenchymal transition (EMT) is closely implicated in the pathogenesis of idiopathic pulmonary fibrosis. Associated with this phenotypic transition is the acquisition of an elongated cell morphology and establishment of stress fibers. The extent to which these EMT-associated changes influence cellular mechanics is unclear. We assessed the biomechanical properties of alveolar epithelial cells (A549) following exposure to TGF-beta 1. Using atomic force microscopy, changes in cell stiffness and surface membrane features were determined. Stimulation with TGF-beta 1 gave rise to a significant increase in stiffness, which was augmented by a collagen I matrix. Additionally, TGF-beta 1-treated cells exhibited a rougher surface profile with notable protrusions. Simultaneous quantitative examination of the morphological attributes of stimulated cells using an image-based high-content analysis system revealed dramatic alterations in cell shape, F-actin content and distribution. Together, these investigations point to a strong correlation between the cytoskeletal-associated cellular architecture and the mechanical dynamics of alveolar epithelial cells undergoing EMT.