Cell locomotion and focal adhesions are regulated by substrate flexibility
Cell locomotion and focal adhesions are regulated by substrate flexibility
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DOI：
10.1073/pnas.94.25.13661
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发表时间：
1997-12-09
影响因子：
11.1
通讯作者：
Wang, YL
中科院分区：
文献类型：
--
作者：
Pelham, RJ;Wang, YL
通过在胶原包被的聚丙烯酰胺底物上培养正常大鼠肾上皮细胞和3T3成纤维细胞，研究了细胞对粘附底物力学性能的反应，这种底物在保持恒定化学环境的同时允许灵活性的变化。与刚性基质上的细胞相比，柔性基质上的细胞扩散减少，运动性或板足活性增加。荧光长春素的显微注射表明，柔性基质上的黏附具有不规则形状和高度动态，而刚性基质上的黏附具有正常形态和稳定得多。在柔性底物上的细胞在粘附位点也含有少量的磷酸酪氨酸。用氧化苯larsin（一种酪氨酸磷酸酶抑制剂）处理这些细胞，诱导形成正常、稳定的局灶粘连，类似于在坚固底物上的粘连。相反，用肌球蛋白抑制剂2,3-丁二酮单肟或KT5926处理坚硬底物上的细胞会导致黏附位点的血管蛋白和磷酸酪氨酸的减少。这些结果表明细胞有能力调查其周围环境的机械特性，并提示在这一过程中可能涉及蛋白酪氨酸磷酸化和肌球蛋白产生的皮质力。这种对物理参数的响应可能代表了复杂生物体中细胞与周围环境相互作用的重要机制。
Responses of cells to mechanical properties of the adhesion substrate were examined by culturing normal rat kidney epithelial and 3T3 fibroblastic cells on a collagen-coated polyacrylamide substrate that allows the flexibility to be varied while maintaining a constant chemical environment. Compared with cells on rigid substrates, those on flexible substrates showed reduced spreading and increased rates of motility or lamellipodial activity. Microinjection of fluorescent vinculin indicated that focal adhesions on flexible substrates were irregularly shaped and highly dynamic whereas those on firm substrates had a normal morphology and were much more stable. Cells on flexible substrates also contained a reduced amount of phosphotyrosine at adhesion sites. Treatment of these cells with phenylarsine oxide, a tyrosine phosphatase inhibitor, induced the formation of normal, stable focal adhesions similar to those on firm substrates. Conversely, treatment of cells on firm substrates with myosin inhibitors 2,3-butanedione monoxime or KT5926 caused the reduction of both vinculin and phosphotyrosine at adhesion sites. These results demonstrate the ability of cells to survey the mechanical properties of their surrounding environment and suggest the possible Involvement of both protein tyrosine phosphorylation and myosin-generated cortical forces In this process. Such response to physical parameters likely represents an important mechanism of cellular interaction with the surrounding environment within a complex organism.