Cell cycle-dependent force transmission in cancer cells.
Cell cycle-dependent force transmission in cancer cells.
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DOI：
10.1091/mbc.e17-12-0726
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发表时间：
2018-10-15
影响因子：
3.3
通讯作者：
Ferrari A
中科院分区：
文献类型：
--
作者：
Panagiotakopoulou M;Lendenmann T;Pramotton FM;Giampietro C;Stefopoulos G;Poulikakos D;Ferrari A
牵引力的产生及其向细胞外环境的传递支持细胞从原发性肿瘤的播散性迁移。在癌细胞中，细胞周期中核刚度的周期性变化提供了有效易位和增殖之间的功能联系。然而，完成这幅图画的机械框架仍未被探索。在这里，Fucci2 报告基因在各种人类上皮癌细胞中表达，以解决其细胞周期相变。最近开发的无参考共焦牵引力显微镜平台捕获了相应的牵引力。组合方法有利于分析个体整合素接触水平上的相依赖性力变化。在 G1 和早期 S 期检测到的力总是高于随后的 S/G2 晚期，并且局部与高水平的桩蛋白磷酸化共定位。通过粘着斑激酶 (FAK) 的生化抑制或转染不可磷酸化或拟磷酸化桩蛋白突变体获得的粘着斑处桩蛋白磷酸化的扰动显着减弱了传递至底物的力。这些数据表明，癌细胞的细胞周期进展过程中力传递的可重复调节，有助于它们入侵密集环境。此外，他们描绘了一个模型，其中桩蛋白磷酸化支持将力传递到基质的粘附的机械成熟。
The generation of traction forces and their transmission to the extracellular environment supports the disseminative migration of cells from a primary tumor. In cancer cells, the periodic variation of nuclear stiffness during the cell cycle provides a functional link between efficient translocation and proliferation. However, the mechanical framework completing this picture remains unexplored. Here, the Fucci2 reporter was expressed in various human epithelial cancer cells to resolve their cell cycle phase transition. The corresponding tractions were captured by a recently developed reference-free confocal traction-force microscopy platform. The combined approach was conducive to the analysis of phase-dependent force variation at the level of individual integrin contacts. Detected forces were invariably higher in the G1 and early S phases than in the ensuing late S/G2, and locally colocalized with high levels of paxillin phosphorylation. Perturbation of paxillin phosphorylation at focal adhesions, obtained through the biochemical inhibition of focal adhesion kinase (FAK) or the transfection of nonphosphorylatable or phosphomimetic paxillin mutants, significantly diminished the force transmitted to the substrate. These data demonstrate a reproducible modulation of force transmission during the cell cycle progression of cancer cells, instrumental to their invasion of dense environments. In addition, they delineate a model in which paxillin phosphorylation supports the mechanical maturation of adhesions relaying forces to the substrate.