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In epithelia, breakdown of tensional homeostasis is closely associated with E-cadherin dysfunction and disruption of tissue function and integrity. In this study, we investigated the effect of E-cadherin mutations affecting distinct protein domains on tensional homeostasis of gastric cancer cells. We used micropattern traction microscopy to measure temporal fluctuations of cellular traction forces in AGS cells transfected with the wild-type E-cadherin or with variants affecting the extracellular, the juxtamembrane, and the intracellular domains of the protein. We focused on the dynamic aspect of tensional homeostasis, namely the ability of cells to maintain a consistent level of tension, with low temporal variability around a set point. Cells were cultured on hydrogels micropatterned with different extracellular matrix (ECM) proteins to test whether the ECM adhesion impacts cell behavior. A combination of Fibronectin and Vitronectin was used as a substrate that promotes the adhesive ability of E-cadherin dysfunctional cells, whereas Collagen VI was used to test an unfavorable ECM condition. Our results showed that mutations affecting distinct E-cadherin domains influenced differently cell tensional homeostasis, and pinpointed the juxtamembrane and intracellular regions of E-cadherin as the key players in this process. Furthermore, Fibronectin and Vitronectin might modulate cancer cell behavior towards tensional homeostasis.
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Effect of correlation between traction forces on tensional homeostasis in clusters of endothelial cells and fibroblasts
The ability of cells to maintain a constant level of cytoskeletal tension in response to external and internal disturbances is referred to as tensional homeostasis. It is essential for the normal physiological function of cells and tissues, and for protection against disease progression, including atherosclerosis and cancer. In previous studies, we defined tensional homeostasis as the ability of cells to maintain a consistent level of cytoskeletal tension with low temporal fluctuations. In those studies, we measured temporal fluctuations of cell-substrate traction forces in clusters of endothelial cells and of fibroblasts. We observed those temporal fluctuations to decrease with increasing cluster size in endothelial cells, but not in fibroblasts. We quantified temporal fluctuation, and thus homeostasis, through the coefficient of variation (CV) of the traction field; the lower the value of CV, the closer the cell is to the state of tensional homeostasis. This metric depends on correlation between individual traction forces. In this study, we analyzed the contribution of correlation between traction forces on traction field CV in clusters of endothelial cells and fibroblasts using experimental data that we had obtained previously. Results of our analysis showed that positive correlation between traction forces was detrimental to homeostasis, and that it was cell type-dependent.
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- PAR ID:
- 10155254
- Date Published:
- Journal Name:
- Journal of biomechanics
- Volume:
- 100
- ISSN:
- 0021-9290
- Page Range / eLocation ID:
- 109588
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/https://doi.org/10.1016/j.jbiomech.2019.109588
