Dissipation monitoring for assessing EGF-induced changes of cell adhesion

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.

► Changes in energy dissipation factor are correlated with changes in level of focal adhesions. ► EGF induces a dynamic restructuring of focal adhesions. ► EGF-induced restructuring of focal adhesions is mediated temporally by EGFR signaling pathways.