Head and neck squamous cell carcinoma (HNSCC) is a worldwide leading malignancy with poor prognoses. Aggressive HNSCC is manifested by forming tumor satellites in the invasive front, which is closely associated with epithelial-mesenchymal transition, local invasion, and metastasis. Limited by the pathological analyses of static cancer specimens conducted in most previous investigations, the dynamic processes and the decisive factors of tumor satellite formation in HNSCC cannot be monitored and studied. The establishment of a system to recapitulate the phenomenon in vitro may be instrumental to explicitly address the question. In this study, we explored the feasibility of establishing an in vitro system to induce tumor satellite formation of different HNSCC cells by applying a system composed of three-dimensional collagen scaffolds. The real-time dynamic process of tumor satellite formation could be monitored in detail, and the pivotal factors accounting for HNSCC tumor satellite formation were evaluated. E-cadherin remodeling, vimentin aggregation, invadopodia formation, and extracellular matrix degradation occurred in the HNSCC cells when they were cultured in an environment created with a low calcium concentration and steric collagen scaffolds, which altogether contributed to tumor satellite formation and spreading. The phenomena of HNSCC cell transition observed in the current system were confirmed in vivo with compatible findings in surgical specimens. Through the use of this system, we illustrated a novel method to study tumor satellite formation in HNSCC. It may serve as a platform for further investigation of underlying mechanisms for tumor satellites of cancer.

头颈部鳞状细胞癌（HNSCC）是全球领先的恶性肿瘤，预后较差。侵袭性HNSCC表现为在侵袭性前沿形成肿瘤卫星，这与上皮-间质转化，局部侵袭和转移密切相关。由于以往大多数研究对静态癌标本进行的病理分析的局限性，无法监测和研究HNSCC中肿瘤卫星形成的动态过程和决定性因素。建立在体外概括现象的系统可能有助于明确解决该问题。在这项研究中，我们探索了建立体外的可行性通过应用由三维胶原蛋白支架组成的系统来诱导不同HNSCC细胞的肿瘤卫星形成的系统。可以详细监测肿瘤卫星形成的实时动态过程，并对影响HNSCC肿瘤卫星形成的关键因素进行了评估。当在低钙浓度和空间胶原支架形成的环境中培养HNSCC细胞时，E-钙黏着蛋白重塑，波形蛋白聚集，内脏足形成和细胞外基质降解发生，这完全有助于肿瘤卫星的形成和扩散。在体内证实了在当前系统中观察到的HNSCC细胞过渡现象在手术标本中具有可比的发现。通过使用该系统，我们说明了一种研究HNSCC中肿瘤卫星形成的新方法。它可以作为进一步研究癌症肿瘤卫星潜在机制的平台。