Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
FASEB J. 2010 Jun;24(6):1838-51. doi: 10.1096/fj.09-151639. Epub 2010 Jan 22.
Vimentin is used widely as a marker of the epithelial to mesenchymal transitions (EMTs) that take place during embryogenesis and metastasis, yet the functional implications of the expression of this type III intermediate filament (IF) protein are poorly understood. Using form factor analysis and quantitative Western blotting of normal, metastatic, and vimentin-null cell lines, we show that the level of expression of vimentin IFs (VIFs) correlates with mesenchymal cell shape and motile behavior. The reorganization of VIFs caused by expressing a dominant-negative mutant or by silencing vimentin with shRNA (neither of which alter microtubule or microfilament assembly) causes mesenchymal cells to adopt epithelial shapes. Following the microinjection of vimentin or transfection with vimentin cDNA, epithelial cells rapidly adopt mesenchymal shapes coincident with VIF assembly. These shape transitions are accompanied by a loss of desmosomal contacts, an increase in cell motility, and a significant increase in focal adhesion dynamics. Our results demonstrate that VIFs play a predominant role in the changes in shape, adhesion, and motility that occur during the EMT.
波形蛋白被广泛用作上皮细胞到间充质转化 (EMT) 的标志物,这种转化发生在胚胎发生和转移过程中,但这种 III 型中间丝 (IF) 蛋白表达的功能意义还知之甚少。通过对正常、转移和波形蛋白缺失细胞系的形态因子分析和定量 Western 印迹分析,我们表明波形蛋白 IF (VIF) 的表达水平与间充质细胞的形状和运动行为相关。表达显性负突变体或用 shRNA 沉默波形蛋白 (这两种方法都不改变微管或微丝组装) 导致 VIF 的重排,使间充质细胞呈现上皮细胞形状。在微注射波形蛋白或转染波形蛋白 cDNA 后,上皮细胞迅速呈现间充质形状,同时 VIF 组装。这些形状转变伴随着桥粒连接的丧失、细胞迁移性的增加和粘着斑动力学的显著增加。我们的结果表明,VIF 在 EMT 过程中发生的形状、黏附和迁移变化中起主要作用。