Department of Biological Physics, Eotvos Lorand University, Budapest, Hungary.
Department of Biochemistry, Eotvos Lorand University, Budapest, Hungary.
PLoS Comput Biol. 2019 Oct 25;15(10):e1007431. doi: 10.1371/journal.pcbi.1007431. eCollection 2019 Oct.
Non-muscle myosin II (NMII)-induced multicellular contractility is essential for development, maintenance and remodeling of tissue morphologies. Dysregulation of the cytoskeleton can lead to birth defects or enable cancer progression. We demonstrate that the Matrigel patterning assay, widely used to characterize endothelial cells, is a highly sensitive tool to evaluate cell contractility within a soft extracellular matrix (ECM) environment. We propose a computational model to explore how cell-exerted contractile forces can tear up the cell-Matrigel composite material and gradually remodel it into a network structure. We identify measures that are characteristic for cellular contractility and can be obtained from image analysis of the recorded patterning process. The assay was calibrated by inhibition of NMII activity in A431 epithelial carcinoma cells either directly with blebbistatin or indirectly with Y27632 Rho kinase inhibitor. Using Matrigel patterning as a bioassay, we provide the first functional demonstration that overexpression of S100A4, a calcium-binding protein that is frequently overexpressed in metastatic tumors and inhibits NMIIA activity by inducing filament disassembly, effectively reduces cell contractility.
非肌肉肌球蛋白 II（NMII）诱导的多细胞收缩对于组织形态的发育、维持和重塑至关重要。细胞骨架的失调可导致出生缺陷或促进癌症进展。我们证明，广泛用于表征内皮细胞的 Matrigel 图案形成测定法是一种高度灵敏的工具，可用于评估软细胞外基质（ECM）环境中细胞的收缩性。我们提出了一个计算模型来探索细胞施加的收缩力如何撕裂细胞-Matrigel 复合材料，并逐渐将其重塑为网络结构。我们确定了一些特征性的细胞收缩性措施，可以通过记录的图案形成过程的图像分析获得。该测定法通过直接用 blebbistatin 或间接用 Y27632 Rho 激酶抑制剂抑制 A431 上皮癌细胞中的 NMII 活性进行校准。我们使用 Matrigel 图案形成作为生物测定法，首次提供了功能证明，即过度表达 S100A4（一种钙结合蛋白，在转移性肿瘤中经常过度表达，并通过诱导纤维丝解聚抑制 NMIIA 活性）可有效降低细胞收缩性。
