Ewing sarcoma (ES) is an aggressive bone and soft-tissue pediatric cancer. High vitronectin (VN) expression has been associated with poor prognosis in other cancers, and we aimed to determine the utility of this extracellular matrix glycoprotein as a biomarker of aggressiveness in ES. Silk fibroin plus gelatin–tyramine hydrogels (HGs) were fabricated with and without cross-linked VN and cultivated with A673 and PDX73 ES cell lines for two and three weeks. VN secretion to culture media was assessed using ELISA. Morphometric analysis was applied for phenotypic characterization. VN release to culture media was higher in 3D models than in monolayer cultures, and intracellular, intercellular, and pericluster presence was also observed. A673-HGs showed lower density of clusters but a proportion of larger clusters than PDX73-HGs, which presented low cluster circularity. The cluster density of A673-HGs without added VN was higher than with added VN and slightly lower in the case of PDX73-HGs. Furthermore, a culture time of three weeks provided no benefits in cluster growth compared to two weeks, especially in A673-HGs. These advances in 3D modeling and digital quantification pave the way for future studies in ES and other cancers to deepen understanding about intra- and intercellular heterogeneity and anti-adhesion VN therapies.
尤文肉瘤（ES）是一种侵袭性骨与软组织儿童恶性肿瘤。高玻连蛋白（VN）表达在其他癌症中与不良预后相关，本研究旨在评估这种细胞外基质糖蛋白作为ES侵袭性生物标志物的应用价值。研究制备了含或不含交联VN的丝素蛋白-明胶-酪胺水凝胶（HGs），分别与A673和PDX73两种ES细胞系进行为期两周和三周的培养。通过ELISA检测培养基中VN分泌水平，并采用形态计量学分析进行表型表征。结果显示，三维模型培养基中VN释放量高于单层培养，同时观察到VN在细胞内、细胞间及细胞团周围的分布。A673-HGs细胞团密度低于PDX73-HGs，但大尺寸细胞团比例更高，而PDX73-HGs细胞团圆形度较低。未添加VN的A673-HGs细胞团密度高于添加VN组，PDX73-HGs则呈现轻微下降趋势。此外，与两周培养相比，三周培养并未促进细胞团生长，在A673-HGs中尤为明显。这些三维建模与数字化定量分析技术的进展，为未来深入研究ES及其他癌症的细胞内外异质性以及抗黏附VN疗法奠定了基础。
肿瘤(癌症)患者之家