Xu Han, Bunde Katie A, Figueiredo Joana, Seruca Raquel, Smith Michael L, Stamenović Dimitrije
Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
Epithelial Interactions in Cancer, Instituto de Investigação e Inovação em Saúde (i3S), University of Porto, 4200-135 Porto, Portugal.
Cancers (Basel). 2022 May 29;14(11):2690. doi: 10.3390/cancers14112690.
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.
在上皮组织中，张力稳态的破坏与E-钙黏蛋白功能障碍以及组织功能和完整性的破坏密切相关。在本研究中，我们调查了影响不同蛋白结构域的E-钙黏蛋白突变对胃癌细胞张力稳态的影响。我们使用微图案牵引力显微镜来测量转染了野生型E-钙黏蛋白或影响该蛋白细胞外、近膜和细胞内结构域的变体的AGS细胞中细胞牵引力的时间波动。我们关注张力稳态的动态方面，即细胞维持一致张力水平的能力，在设定点周围具有较低的时间变异性。将细胞培养在微图案化有不同细胞外基质（ECM）蛋白的水凝胶上，以测试ECM黏附是否影响细胞行为。使用纤连蛋白和玻连蛋白的组合作为促进E-钙黏蛋白功能失调细胞黏附能力的底物，而使用VI型胶原蛋白来测试不利的ECM条件。我们的结果表明，影响不同E-钙黏蛋白结构域的突变对细胞张力稳态的影响不同，并确定E-钙黏蛋白的近膜和细胞内区域是这一过程中的关键因素。此外，纤连蛋白和玻连蛋白可能调节癌细胞对张力稳态的行为。
