Dissipation monitoring for assessing EGF-induced changes of cell adhesion
Dissipation monitoring for assessing EGF-induced changes of cell adhesion
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DOI：
10.1016/j.bios.2012.06.018
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发表时间：
2012-10-01
影响因子：
12.6
通讯作者：
Xi, Jun
中科院分区：
文献类型：
--
作者：
Chen, Jennifer Y.;Shahid, Ammar;Xi, Jun
表皮生长因子（EGF）诱导的细胞去黏附被认为是正常胚胎发育、伤口修复、炎症反应以及肿瘤细胞转移的关键步骤。像许多其他细胞过程一样，这种细胞去黏附呈现出一种复杂的、具有时间依赖性的模式。对这一过程的全面理解需要在分子水平上对细胞 - 基质相互作用进行定量的实时评估。我们利用耗散型石英晶体微天平（QCM - D）成功地实时追踪了单层MCF10A细胞在表皮生长因子诱导下能量耗散因子ΔD的变化。这种具有时间依赖性的ΔD响应在定性和定量上都与细胞在表皮生长因子作用下黏着斑快速解体、转变以及缓慢重新组装的连续事件密切相关。基于这种强相关性，我们利用QCM - D证明这种动态的黏着斑重构在时间上受到表皮生长因子受体（EGFR）信号传导的下游通路，如磷脂酰肌醇 - 3 - 激酶（PI3K）、丝裂原活化蛋白激酶/细胞外调节蛋白激酶（MAPK/ERK）和磷脂酶C（PLC）通路的调控。由于QCM - D是一种非侵入性技术，这种新方法在细胞过程的基础研究中可能具有广泛的应用，例如细胞信号传导、运输以及机械力转导，并且在药物和生物标志物筛选方面具有应用前景。（C）2012爱思唯尔公司。版权所有。
Epidermal growth factor (EGF)-induced cell de-adhesion has been implicated as a critical step of normal embryonic development, wound repair, inflammatory response, and tumor cell metastasis. Like many other cellular processes, this cell de-adhesion exhibits a complex, time-dependent pattern. A comprehensive understanding of this process requires a quantitative, real-time assessment of cell-substrate interactions at the molecular level. We employed the quartz crystal microbalance with dissipation monitoring (QCM-D) to successfully track the EGF-induced changes in energy dissipation factor, Delta D, of a monolayer of MCF10A cells in real time. This time-dependent Delta D response correlates well both qualitatively and quantitatively with sequential events of a rapid disassembly, transition, and slow reassembly of focal adhesions of the cells in response to EGF exposure. Based on this strong correlation, we utilized the QCM-D to demonstrate that this dynamic focal-adhesion restructuring is regulated temporally by the downstream pathways of EGFR signaling such as the PI3K, MAPK/ERK, and PLC pathways. Because the QCM-D is a noninvasive technique, this novel approach potentially has a broad range of applications in the fundamental study of cellular processes, such as cell signaling and trafficking and mechanotransduction, and holds promise for drug and biomarker screening. (C) 2012 Elsevier B.V. All rights reserved.