Rearrangements of desmosomal and cytoskeletal proteins during the transition from epithelial to fibroblastoid organization in cultured rat bladder carcinoma cells.
Rearrangements of desmosomal and cytoskeletal proteins during the transition from epithelial to fibroblastoid organization in cultured rat bladder carcinoma cells.
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DOI：
10.1083/jcb.109.4.1495
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发表时间：
1989-10
期刊：
影响因子：
--
通讯作者：
Thiery JP
中科院分区：
文献类型：
--
作者：
Boyer B;Tucker GC;Vallés AM;Franke WW;Thiery JP
已知细胞形态和分化状态的变化在胚胎发生和癌发生中起重要作用。特别深刻的变化的例子是上皮细胞向间充质细胞的转化;即，部分或全部多角形极性上皮细胞的解离，并转化为高运动性的细长成纤维细胞样细胞。作为这种细胞形态学变化的体外模型系统，我们使用大鼠膀胱癌衍生细胞系NBT-Ⅱ的细胞培养物，该细胞系在暴露于含有商业血清替代物的诱导培养基时，（Ultroser G），显示其组织的广泛变化（上皮-间充质转化）：上皮细胞之间的连接分裂，上皮细胞组织丧失，并且所产生的单个细胞变得能动并呈现梭形成纤维细胞样外观。使用免疫荧光显微镜和生化蛋白质表征技术，我们表明，这种变化是伴随着桥粒斑块蛋白（桥粒斑蛋白，桥粒芯蛋白，斑珠蛋白）的重新分配和重组的细胞角蛋白和肌动蛋白胞衬蛋白丝系统。此外，在成纤维细胞样细胞中形成波形蛋白型的中等大小的细丝。我们证明了桥粒蛋白的调节，特别是可溶性桥粒斑蛋白的增加，是细胞解离和上皮-间充质转化中相对早期的事件。在这个过程中，5小时的潜伏期后，诱导培养基的加入之前，这些桥粒组分从质膜上除去。这种可逆的转变依赖于持续的蛋白质合成和磷酸化，但不依赖于诱导介质的存在，而不依赖于最初的2小时周期。我们讨论了这个实验系统的价值作为一个生理相关的方法，用于研究桥粒和其他细胞间粘附结构的组装和拆卸的调节，并作为一个模型的细胞从一个分化状态转换成另一个。
Changes of cell morphology and the state of differentiation are known to play important roles in embryogenesis as well as in carcinogenesis. Examples of particularly profound changes are the conversions of epithelial to mesenchymal cells; i.e., the dissociation of some or all polygonal, polar epithelial cells and their transformation into elongate, fibroblastoid cells of high motility. As an in vitro model system for such changes in cell morphology, we have used cell cultures of the rat bladder carcinoma-derived cell line NBT-II which, on exposure to inducing medium containing a commercial serum substitute (Ultroser G), show an extensive change in their organization (epithelial-mesenchymal transition): the junctions between the epithelial cells are split, the epithelial cell organization is lost, and the resulting individual cells become motile and assume a spindle- like fibroblastoid appearance. Using immunofluorescence microscopy and biochemical protein characterization techniques, we show that this change is accompanied by a redistribution of desmosomal plaque proteins (desmoplakins, desmoglein, plakoglobin) and by a reorganization of the cytokeratin and the actin-fodrin filament systems. Moreover, intermediate-sized filaments of the vimentin type are formed in the fibroblastoid cells. We demonstrate that the modulation of desmosomal proteins, specifically an increase in soluble desmoplakins, is a relatively early event in cell dissociation and in epithelial- mesenchymal transition. In this process, a latent period of 5 h upon addition of inducing medium precedes the removal of these desmosomal components from the plasma membrane. The transition, which is reversible, is dependent on continued protein synthesis and phosphorylation but not on the presence of the inducing medium beyond the initial 2-h period. We discuss the value of this experimental system as a physiologically relevant approach for studying the regulation of the assembly and disassembly of desmosomes and other intercellular adhesion structures, and as a model of the conversion of cells from one state of differentiation into another.