Dept. of Cell Pathology, Faculty of Biotechnology, University of Wroclaw, Poland.
Eur J Histochem. 2010 Apr 13;54(2):e14. doi: 10.4081/ejh.2010.e14.
The dynamic reorganization of the actin cytoskeleton is regulated by a number of actin binding proteins (ABPs). Four human colon adenocarcinoma cell lines - parental and three selected sublines, which differ in motility and metastatic potential, were used to investigate the expression level and subcellular localization of selected ABPs. Our interest was focused on cofilin and ezrin. These proteins are essential for cell migration and adhesion. The data received for the three more motile adenocarcinoma sublines (EB3, 3LNLN, 5W) were compared with those obtained for the parental LS180 adenocarcinoma cells and fibroblastic NRK cells. Quantitative densitometric analysis and confocal fluorescence microscopy were used to examine the expression levels and subcellular distribution of the selected ABPs. Our data show distinct increase in the level of cofilin in adenocarcinoma cells accompanied by the reduction of inactive phosphorylated form of cofilin. In more motile cells, cofilin was accumulated at cellular periphery in co-localization with actin filaments. Furthemore, we indicated translocation of ezrin towards the cell periphery within more motile cells in comparison with NRK and parental adenocarcinoma cells. In summary, our data indicate the correlation between migration ability of selected human colon adenocarcinoma sublines and subcellular distribution as well as the level of cofilin and ezrin. Therefore these proteins might be essential for the higher migratory activity of invasive tumor cells.
细胞骨架的动态重组受许多肌动蛋白结合蛋白(ABPs)的调节。我们使用了四种人结肠腺癌细胞系——亲本细胞系和三个在运动性和转移潜能上不同的选定亚系,来研究选定的 ABPs 的表达水平和亚细胞定位。我们的兴趣集中在丝切蛋白和埃兹蛋白上。这些蛋白质对细胞迁移和黏附至关重要。我们将三个更具运动性的腺癌亚系(EB3、3LNLN、5W)的数据与亲本 LS180 腺癌细胞和成纤维细胞 NRK 细胞获得的数据进行了比较。定量密度分析和共聚焦荧光显微镜用于检查选定 ABPs 的表达水平和亚细胞分布。我们的数据表明,在腺癌细胞中,丝切蛋白的水平明显增加,同时无活性磷酸化形式的丝切蛋白减少。在更具运动性的细胞中,丝切蛋白在细胞边缘与肌动蛋白丝共定位聚集。此外,与 NRK 和亲本腺癌细胞相比,我们发现埃兹蛋白在更具运动性的细胞中向细胞边缘易位。总之,我们的数据表明,所选人结肠腺癌亚系的迁移能力与丝切蛋白和埃兹蛋白的亚细胞分布和水平之间存在相关性。因此,这些蛋白质可能对侵袭性肿瘤细胞更高的迁移活性至关重要。