Loss of giant obscurins alters breast epithelial cell mechanosensing of matrix stiffness.
Loss of giant obscurins alters breast epithelial cell mechanosensing of matrix stiffness.
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DOI：
10.18632/oncotarget.10997
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发表时间：
2017-08-15
期刊：
影响因子：
--
通讯作者：
Konstantopoulos K
中科院分区：
文献类型：
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作者：
Stroka KM;Wong BS;Shriver M;Phillip JM;Wirtz D;Kontrogianni-Konstantopoulos A;Konstantopoulos K
Obscurins是一个含有RhoGEF的蛋白家族，在乳腺上皮中具有肿瘤和转移抑制作用。在正常乳腺上皮细胞中下调巨暗蛋白导致活性RhoA及其下游效应物水平降低。在此，我们阐明了如何耗尽的巨大obscuris影响乳腺上皮细胞的微环境的机械性能的变化的反应。我们发现，敲低obscurins增加细胞形态动力学，迁移速度，并在聚丙烯酰胺凝胶≥ 1 kPa的扩散，大概是通过减少粘着斑面积和密度以及细胞牵引力。暗蛋白的耗尽也增加了细胞在软（0.4- 4kPa）表面上的机械敏感性。与obscurin下调相似，乳腺上皮细胞中Rho激酶的药理学抑制增加了迁移和形态动力学，表明obscurin敲低后Rho激酶活性的抑制可以解释形态动力学和迁移的改变。相反，肌球蛋白轻链激酶的抑制减少形态动力学和迁移，表明细胞形状的时间变化是有效迁移所必需的。总的来说，下调巨暗蛋白通过改变细胞机械生物学促进细胞迁移通过不同刚度的异质微环境。
Obscurins are a family of RhoGEF-containing proteins with tumor and metastasis suppressing roles in breast epithelium. Downregulation of giant obscurins in normal breast epithelial cells leads to reduced levels of active RhoA and of its downstream effectors. Herein, we elucidate how depletion of giant obscurins affects the response of breast epithelial cells to changes in the mechanical properties of the microenvironment. We find that knockdown of obscurins increases cell morphodynamics, migration speed, and diffusivity on polyacrylamide gels of ≥ 1 kPa, presumably by decreasing focal adhesion area and density as well as cell traction forces. Depletion of obscurins also increases cell mechanosensitivity on soft (0.4–4 kPa) surfaces. Similar to downregulation of obscurins, pharmacological inhibition of Rho kinase in breast epithelial cells increases migration and morphodynamics, suggesting that suppression of Rho kinase activity following obscurin knockdown can account for alterations in morphodynamics and migration. In contrast, inhibition of myosin light chain kinase reduces morphodynamics and migration, suggesting that temporal changes in cell shape are required for efficient migration. Collectively, downregulation of giant obscurins facilitates cell migration through heterogeneous microenvironments of varying stiffness by altering cell mechanobiology.