Department of Chemistry, Drexel University, 3141 Chestnut Street, Philadelphia, PA 19104, USA.
Biosens Bioelectron. 2012 Oct-Dec;38(1):375-81. doi: 10.1016/j.bios.2012.06.018. Epub 2012 Jun 19.
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, ΔD, of a monolayer of MCF10A cells in real time. This time-dependent ΔD 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.
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、炎症反应和肿瘤细胞转移的关键步骤。像许多其他细胞过程一样，这种细胞去黏附表现出复杂的、时变的模式。要全面了解这一过程，需要在分子水平上对细胞-基底相互作用进行定量、实时评估。我们采用石英晶体微天平（QCM-D）成功地实时跟踪 MCF10A 细胞单层中 EGF 诱导的能量耗散因子 ΔD 的变化。这种时变的 ΔD 响应与细胞对 EGF 暴露的快速解组装、过渡和缓慢再组装的焦点黏附的顺序事件在定性和定量上都很好地相关。基于这种强相关性，我们利用 QCM-D 证明了这种动态焦点黏附重构是由 EGFR 信号的下游途径（如 PI3K、MAPK/ERK 和 PLC 途径）在时间上调节的。由于 QCM-D 是一种非侵入性技术，这种新方法在细胞信号转导、细胞迁移和力学转导等细胞过程的基础研究中具有广泛的应用前景，并有望用于药物和生物标志物筛选。
