Department of Cell Biology, Faculty of Sciences, University of Utrecht, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
Cell Mol Life Sci. 2013 Jan;70(2):293-307. doi: 10.1007/s00018-012-1130-2. Epub 2012 Aug 25.
Integrin-mediated cytoskeletal tension supports growth-factor-induced proliferation, and disruption of the actin cytoskeleton in growth factor-stimulated cells prevents the re-expression of cyclin D and cell cycle re-entry from quiescence. In contrast to cells that enter the cell cycle from G0, cycling cells continuously express cyclin D, and are subject to major cell shape changes during the cell cycle. Here, we investigated the cell cycle requirements for cytoskeletal tension and cell spreading in cycling mammalian cells that enter G1-phase from mitosis. Disruption of the actin cytoskeleton at progressive time-points in G1-phase induced cell rounding, FA disassembly, and attenuated both integrin signaling and growth factor-induced p44/p42 mitogen-activated protein kinase activation. Although cyclin D expression was reduced, the expression of cyclin A and entry into S-phase were not affected. Moreover, expression of cyclin B1, progression through G2- and M-phase, and commitment to a new cell cycle occurred normally. In contrast, cell cycle progression was strongly prevented by inhibition of MAPK activity in G1-phase, whereas cell spreading, cytoskeletal organization, and integrin signaling were not impaired. MAPK inhibition also prevented cytoskeleton-independent cell cycle progression. Thus, these results uncouple the requirements for cell spreading and cytoskeletal organization from MAPK signaling, and show that cycling mammalian cells can proliferate independently of actin stress fibers, focal adhesions, or cell spreading, as long as a threshold level of MAPK activity is sustained.
整合素介导的细胞骨架张力支持生长因子诱导的增殖,并且在生长因子刺激的细胞中破坏肌动蛋白细胞骨架会阻止细胞从静止期重新表达细胞周期蛋白 D 并重新进入细胞周期。与从 G0 进入细胞周期的细胞不同,循环细胞持续表达细胞周期蛋白 D,并在细胞周期中经历主要的细胞形状变化。在这里,我们研究了从有丝分裂进入 G1 期的循环哺乳动物细胞中细胞骨架张力和细胞铺展的细胞周期要求。在 G1 期的渐进时间点破坏肌动蛋白细胞骨架会诱导细胞变圆、FA 解聚,并减弱整合素信号和生长因子诱导的 p44/p42 丝裂原激活蛋白激酶激活。尽管细胞周期蛋白 D 的表达减少,但细胞周期蛋白 A 的表达和 S 期的进入不受影响。此外,细胞周期蛋白 B1 的表达、通过 G2 和 M 期的进展以及向新细胞周期的承诺正常发生。相比之下,MAPK 活性在 G1 期的抑制强烈阻止了细胞周期的进展,而细胞铺展、细胞骨架组织和整合素信号不受影响。MAPK 抑制也阻止了细胞骨架独立的细胞周期进展。因此,这些结果将细胞铺展和细胞骨架组织的要求与 MAPK 信号解耦,并表明循环哺乳动物细胞可以在不依赖肌动蛋白应力纤维、焦点粘连或细胞铺展的情况下增殖,只要维持 MAPK 活性的阈值水平。