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首页> 外文期刊>Life sciences >Gelsolin affects the migratory ability of human colon adenocarcinoma and melanoma cells
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Gelsolin affects the migratory ability of human colon adenocarcinoma and melanoma cells

机译：凝溶胶蛋白影响人结肠腺癌和黑色素瘤细胞的迁移能力

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Aims: Formation of different protrusive structures by migrating cells is driven by actin polymerization at the plasma membrane region. Gelsolin is an actin binding protein controlling the length of actin filaments by its severing and capping activity. The main goal of this study was to determine the effect of gelsolin expression on the migration of human colon adenocarcinoma LS180 and melanoma A375 cells. Main methods: Colon adenocarcinoma cell line LS180 was stably transfected with plasmid containing human cytoplasmic gelsolin cDNA tagged to enhanced green fluorescence protein (EGFP). Melanoma A375 cells were transfected with siRNAs directed against gelsolin. Real-time PCR and Western blotting were used to determine the level of gelsolin. The ability of actin to inhibit DNase I activity was used to quantify monomeric and total actin level and calculate the state of actin polymerization. Fluorescence confocal microscopy was applied to observe gelsolin and vinculin distribution along with actin cytoskeleton organization. Key findings: Increased level of gelsolin expression leads to its accumulation at the submembranous region of the cell accompanied by distinct changes in the state of actin polymerization and an increase in the migration of LS180 cells. In addition, LS180 cells overexpressing gelsolin form podosome-like structures as indicated by vinculin redistribution and its colocalization with gelsolin and actin. Downregulation of gelsolin expression in melanoma A375 cells significantly reduces their migratory potential. Significance: Our experimental data indicate that alterations in the expression level of gelsolin and its subcellular distribution may be directly responsible for determining migration capacity of human cancer cells.
机译：目的：细胞质膜区域的肌动蛋白聚合反应可促使细胞迁移形成不同的突起结构。凝溶胶蛋白是一种肌动蛋白结合蛋白，通过其切断和封闭活性来控制肌动蛋白丝的长度。这项研究的主要目的是确定凝溶胶蛋白表达对人结肠腺癌LS180和黑素瘤A375细胞迁移的影响。主要方法：用含有标记有增强型绿色荧光蛋白（EGFP）的人胞质凝溶胶蛋白cDNA的质粒稳定转染LS180结肠癌细胞。用针对凝溶胶蛋白的siRNA转染黑素瘤A375细胞。实时PCR和蛋白质印迹法用于测定凝溶胶蛋白的水平。肌动蛋白抑制DNase I活性的能力用于定量单体和总肌动蛋白水平，并计算肌动蛋白聚合状态。荧光共聚焦显微镜用于观察凝溶胶蛋白和纽蛋白的分布以及肌动蛋白的细胞骨架组织。关键发现：凝溶胶蛋白表达水平的提高导致其在细胞膜下区域的积累，并伴随着肌动蛋白聚合状态的明显变化和LS180细胞迁移的增加。此外，过表达凝溶胶蛋白的LS180细胞会形成荚膜状结构，这是由长春花素重新分布及其与凝溶胶蛋白和肌动蛋白共定位所表明的。黑色素瘤A375细胞中凝溶胶蛋白表达的下调显着降低了其迁移潜力。启示：我们的实验数据表明凝溶胶蛋白的表达水平及其亚细胞分布的改变可能直接决定了人类癌细胞的迁移能力。

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