The influence of GFP-actin expression on the adhesion dynamics of HepG2 cells on a model extracellular matrix
The influence of GFP-actin expression on the adhesion dynamics of HepG2 cells on a model extracellular matrix
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DOI：
10.1016/j.biomaterials.2005.01.069
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发表时间：
2005-09-01
期刊：
影响因子：
14
通讯作者：
Chan, V
中科院分区：
文献类型：
--
作者：
Feng, ZQ;Chen, WN;Chan, V
整合素是一类重要的细胞表面受体，介导大多数锚定依赖性细胞与天然细胞外基质（ECM）和生物材料的粘附。已知整合素与ECM蛋白的结合触发细胞骨架的机械化学反应。迄今为止，整合素-ECM相互作用和细胞骨架动力学之间的复杂相互作用导致生物材料上的细胞形态发生的调节在很大程度上仍然未知。在这项研究中，绿色荧光蛋白（GFP）-肌动蛋白在HepG 2细胞中表达的时间可视化的细胞骨架结构的贴壁细胞上的天然来源的材料。通过结合共聚焦反射对比显微镜和荧光显微镜，粘附接触动力学，细胞骨架重塑和二维扩展的完整和GFP-肌动蛋白表达的HepG 2细胞油胶原蛋白和纤连蛋白包被的基板同时探测在初始细胞接种。首先，我们的研究结果表明，整合素-胶原或整合素-纤连蛋白相互作用后HepG 2细胞粘附接触的演变受到GFP-肌动蛋白表达的损害。此外，在GFP-肌动蛋白表达后，纤连蛋白和胶原蛋白上细胞变形的初始速率分别降低了70%和43%。有趣的是，HepG 2细胞的稳态粘附能保持不变，并增加纤连蛋白和胶原蛋白包被的基板上，分别在GFP-肌动蛋白表达。我们的高度集成的生物物理方法表明，GFP-肌动蛋白扩散集中在细胞质皮层在最初的细胞接种，而粘附接触的发展和细胞扩散。粘附接触形成的动力学分析表明，细胞骨架特性和ECM蛋白在细胞粘附中存在复杂的相互作用。(c)2005爱思唯尔有限公司保留所有权利。
Integrins belong to a family of important cell surface receptors which mediate the adhesion of most anchorage-dependent cells to nature extracellular matrix (ECM) and biomaterials. It is known that the binding of integrin with ECM proteins triggers mechanochemical responses of cytoskeleton. To date, the intricate interplay between integrin-ECM interaction and cytoskeleton dynamics leading to the regulation of cell morphogenesis on biomaterials remains largely unknown. In this study, green fluorescence protein (GFP)-actins were expressed in HepG2 cells for the temporal visualization of cytoskeletal structure of adherent cells on naturally derived materials. By combining confocal reflectance contrast microscopy and fluorescence microscopy, the adhesion contact dynamics, cytoskeleton remodeling and two-dimensional spreading of intact and GFP-actin expressing HepG2 cells oil collagen and fibronectin-coated substrates are simultaneously probed during the initial cell seeding. First of all, our results show that the evolution of adhesion contact of HepG2 cells upon integrin-collagen or integrin-fibronectin interaction is impaired by GFP-actin expression. Also, the initial rate of cell deformation is reduced by 70% and 43% on fibronectin and collagen, respectively, upon GFP-actin expression. Interestingly, the steady-state adhesion energy of HepG2 cells remains unchanged and increases on fibronectin- and collagen-coated substrate, respectively, upon GFP-actin expression. Our highly integrated biophysical approach demonstrates that GFP-actins diffusively concentrate in the cytoplasmic cortex during initial cell seeding while adhesion contact evolves and cell spreads. Kinetics analysis on the adhesion contact formation demonstrates the intricate interplay between cytoskeleton property and ECM proteins in cell adhesion. (c) 2005 Elsevier Ltd. All rights reserved.