Cykowska Anna, Hofmann Ulf Krister, Tiwari Aadhya, Kosnopfel Corinna, Riester Rosa, Danalache Marina
Department of Orthopaedic Surgery, University Hospital and Faculty of Medicine, University Hospital of Tübingen, Tübingen, Germany.
Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy.
Front Mol Med. 2023 Apr 20;3:1050487. doi: 10.3389/fmmed.2023.1050487. eCollection 2023.
Malignant melanoma is the most lethal form of skin cancer. Y-box binding protein 1 (YB-1) plays a prominent role in mediating metastatic behavior by promoting epithelial-to-mesenchymal transition (EMT). Migratory melanoma cells exhibit two major migration modes: elongated mesenchymal or rounded amoeboid. Using A375 melanoma cell line and the YB-1 knock-out model, we aimed to elucidate biochemical and biomechanical changes in migration signaling pathways in the context of melanoma metastases. We subjected A375 YB-1 knock-out and parental cells to atomic force microscopy (stiffness determination), immunolabelling, and proteome analysis. We found that YB-1 expressing cells were significantly stiffer compared to the corresponding YB-1 knock-out cell line. Our study demonstrated that the constitutive expression of YB-1 in A375 melanoma cell line appears to be closely related to known biomarkers of epithelial-to-mesenchymal transition, nestin, and vimentin, resulting in a stiffer phenotype, as well as a wide array of proteins involved in RNA, ribosomes, and spliceosomes. YB-1 knock-out resulted in nestin depletion and significantly lower vimentin expression, as well as global upregulation of proteins related to the cytoskeleton and migration. YB-1 knock-out cells demonstrated both morphological features and biochemical drivers of mesenchymal/ameboid migration. Melanoma is a highly plastic, adaptable, and aggressive tumor entity, capable of exhibiting characteristics of different migratory modes.
恶性黑色素瘤是皮肤癌中最致命的一种。Y盒结合蛋白1（YB-1）通过促进上皮-间质转化（EMT）在介导转移行为中发挥重要作用。迁移性黑色素瘤细胞表现出两种主要的迁移模式：细长的间充质样或圆形的阿米巴样。利用A375黑色素瘤细胞系和YB-1基因敲除模型，我们旨在阐明黑色素瘤转移背景下迁移信号通路中的生化和生物力学变化。我们对A375 YB-1基因敲除细胞和亲本细胞进行了原子力显微镜检查（硬度测定）、免疫标记和蛋白质组分析。我们发现，与相应的YB-1基因敲除细胞系相比，表达YB-1的细胞明显更硬。我们的研究表明，A375黑色素瘤细胞系中YB-1的组成性表达似乎与上皮-间质转化的已知生物标志物巢蛋白和波形蛋白密切相关，导致更硬的表型，以及一系列参与RNA、核糖体和剪接体的蛋白质。YB-1基因敲除导致巢蛋白耗竭和波形蛋白表达显著降低，以及与细胞骨架和迁移相关的蛋白质整体上调。YB-1基因敲除细胞表现出间充质/阿米巴样迁移的形态学特征和生化驱动因素。黑色素瘤是一种高度可塑性、适应性和侵袭性的肿瘤实体，能够表现出不同迁移模式的特征。
