Associate Professor University of New Haven West Haven, Connecticut, United States
Introduction: : During development and disease progression, cells experience mechanical and osmatic compression that alter cell volume, which is an essential regulator of cellular homeostasis and diverse cellular functions. However, the interplay of the cell volume and intracellular organization, and the complex mechanics of extracellular matrix (ECM) remains largely unknown. This study investigated the effects of volumetric compression on breast cancer cells MCF-7 behaviors, including cell spreading, adhesion formation, and YAP nuclear translocation. In addition, volumetric compression regulates the dynamics of cell-ECM interactions and irreversible ECM remodeling at both single-cell and multicellular level.
Materials and
Methods: : MCF-7 cells were cultured in DMEM with 10% fetal bovin serum (FBS) and 1% penicillin/streptomycin. Cells were maintained at 37 °C in a humidified incubator with 5% CO2 with medium change every two or three days. Cells were passenged using 0.25 EDTA-Trypsin and passage 3-10 were used for the experiments. To apply volumetric compression, cells were treated with 0%, 2% and 4% PEG 200. The effect of compression were checked at different time points, 4hr, 8hr, 12 hr, and 24hr. Cell viability and proliferation were evaluated and compared. Cell morphology, spreading area, protrusion dynamics, and YAP nuclear translocation were measured and compared.
Results, Conclusions, and Discussions:: We first evaluated the effect of volumetric compression on MCF-7 collective cell migration using wound healing assay. Our results showed MCF-7 were jammed with reduced motility and reduced cell migration rate and wound closure rate. We further studied how different levels of compression affect cell spreading area, circularity, and protrusion dynamics. Our results showed MCF-7 cells experience high cytoskeletal dynamics with increased protrusion, reduced cell spreading area, and reduced YAP nuclear translocation. Future work will focus on the investigation of contractility of MCF-7 under volumetric compression using traction force microscopy (TFM).
Acknowledgements and/or References (Optional):: This project is supported by NSF (2342274).
