Several techniques can be used to monitor cell dynamism after a perturbation. Among these, Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D) offers the great advantage to study the mechanical properties of cells in real-time and with a great sensitivity. Here, we used QCM-D to investigate the effects of two cytoskeleton-targeting agents, cytochalasin D (CytoD) and Y27632, on human MCF-7 cells. Cell adhesion on the sensor surface, crucial for in-flow experiments, was obtained by covalent adsorption of a fibronectin (FN) film, an extracellular matrix (ECM) protein. Direct analysis of MCF-7 cells on FN-coated sensor, shows a specific cellular response that was revealed and quantified by QCM-D after drugs exposure. Notably, upon treatment with Y27632, we observed a two-regime dissipation behavior that we associated with specific modifications of actin filaments and signaling proteins providing a link between biophysical and molecular mechanisms. Overall, this approach opens new opportunities for studying cellular response to mechanical cues in different biological conditions.

Quartz Crystal Microbalance as Cell-Based Biosensor to Detect and Study Cytoskeletal Alterations and Dynamics

De Domenico Stefania;Gaballo Antonio;Arima Valentina
2018

Abstract

Several techniques can be used to monitor cell dynamism after a perturbation. Among these, Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D) offers the great advantage to study the mechanical properties of cells in real-time and with a great sensitivity. Here, we used QCM-D to investigate the effects of two cytoskeleton-targeting agents, cytochalasin D (CytoD) and Y27632, on human MCF-7 cells. Cell adhesion on the sensor surface, crucial for in-flow experiments, was obtained by covalent adsorption of a fibronectin (FN) film, an extracellular matrix (ECM) protein. Direct analysis of MCF-7 cells on FN-coated sensor, shows a specific cellular response that was revealed and quantified by QCM-D after drugs exposure. Notably, upon treatment with Y27632, we observed a two-regime dissipation behavior that we associated with specific modifications of actin filaments and signaling proteins providing a link between biophysical and molecular mechanisms. Overall, this approach opens new opportunities for studying cellular response to mechanical cues in different biological conditions.
2018
actin
cytoskeleton
quartz crystal microbalance
Y27632
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.14243/351470
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact