Department of Biology, Cancer Biology Program, Stanford University, Stanford, CA 94305, USA.
Proc Natl Acad Sci U S A. 2010 Jul 27;107(30):13324-9. doi: 10.1073/pnas.1002662107. Epub 2010 Jun 21.
During normal development and in disease, cohesive tissues undergo rearrangements that require integration of signals from cell adhesions to neighboring cells and to the extracellular matrix (ECM). How a range of cell behaviors is coordinated by these different adhesion complexes is unknown. To analyze epithelial cell motile behavior in response to combinations of cell-ECM and cell-cell adhesion cues, we took a reductionist approach at the single-cell scale by using unique, functionalized micropatterned surfaces comprising alternating stripes of ECM (collagenIV) and adjustable amounts of E-cadherin-Fc (EcadFc). On these surfaces, individual cells spatially segregated integrin- and cadherin-based complexes between collagenIV and EcadFc surfaces, respectively. Cell migration required collagenIV and did not occur on surfaces functionalized with only EcadFc. However, E-cadherin adhesion dampened lamellipodia activity on both collagenIV and EcadFc surfaces and biased the direction of cell migration without affecting the migration rate, all in an EcadFc concentration-dependent manner. Traction force microscopy showed that spatial confinement of integrin-based adhesions to collagenIV stripes induced anisotropic cell traction on collagenIV and migration directional bias. Selective depletion of different pools of alphaE-catenin, an E-cadherin and actin binding protein, identified a membrane-associated pool required for E-cadherin-mediated adhesion and down-regulation of lamellipodia activity and a cytosolic pool that down-regulated the migration rate in an E-cadherin adhesion-independent manner. These results demonstrate that there is crosstalk between E-cadherin- and integrin-based adhesion complexes and that E-cadherin regulates lamellipodia activity and cell migration directionality, but not cell migration rate.
在正常发育和疾病过程中，凝聚组织经历重排，需要整合来自细胞黏附与相邻细胞和细胞外基质 (ECM) 的信号。这些不同的黏附复合物如何协调一系列细胞行为尚不清楚。为了分析上皮细胞对细胞-ECM 和细胞-细胞黏附信号组合的运动行为，我们在单细胞尺度上采取了简化方法，使用独特的、功能化的微图案化表面，这些表面由交替的 ECM（胶原 IV）和可调节量的 E-钙黏蛋白-Fc（EcadFc）条纹组成。在这些表面上，单个细胞分别将整联蛋白和钙黏蛋白为基础的复合物空间分隔在胶原 IV 和 EcadFc 表面之间。细胞迁移需要胶原 IV，而仅在功能化有 EcadFc 的表面上不会发生。然而，E-钙黏蛋白黏附在胶原 IV 和 EcadFc 表面上都减弱了片状伪足的活性，并在不影响迁移率的情况下偏向细胞迁移的方向，所有这些都依赖于 EcadFc 浓度的变化。牵引力显微镜显示，整联蛋白为基础的黏附物在胶原 IV 条纹上的空间限制诱导了细胞在胶原 IV 上的各向异性牵引力和迁移方向的偏向。对不同池的 alphaE-连环蛋白（E-钙黏蛋白和肌动蛋白结合蛋白）的选择性耗竭，确定了膜相关池对于 E-钙黏蛋白介导的黏附和片状伪足活性的下调以及细胞骨架池对于 E-钙黏蛋白黏附非依赖性的迁移率下调是必需的。这些结果表明，E-钙黏蛋白和整联蛋白为基础的黏附复合物之间存在串扰，并且 E-钙黏蛋白调节片状伪足的活性和细胞迁移的方向性，但不调节细胞迁移率。
