Background/Objectives: Cutaneous squamous cell carcinoma (cSCC) is a very common skin malignancy of the head and neck area, with a propensity to spread to local lymph nodes. Epithelial-to-mesenchymal transition (EMT) and cancer-associated fibroblasts (CAFs) play a well-documented role in the progression of the disease. In this study, we developed and characterised multicellular tumour spheroids (MCTS) composed of patient-derived metastatic cSCC cell lines—each exhibiting distinct phenotypes—combined with either dermal- or lymph node-derived fibroblasts. We aimed to investigate how these cellular combinations influence MCTS formation, spatial architecture, and invasive behaviour. We hypothesised that the interplay between different cSCC and fibroblast cell combinations would differentially influence spheroid formation and invasion.Methods: Using live-cell microscopy we assessed the spatial architectures specific to each cell line-fibroblast combination and evaluated the expression of EMT and CAF markers. Furthermore, we utilised MCTS in invasion models to investigate associations between the mode of invasion and the EMT phenotype of the cancer cell line.Results: We show that metastatic cSCC/fibroblast MCTS self-organise into distinct spatial architectures. They also invade through collagen in a manner influenced by fibroblasts but dominated by the EMT status of the originating cancer cells.Conclusions: These findings highlight the physiological relevance and utility of MCTS as models for investigating tumour–stroma interactions and invasion dynamics in metastatic cSCC.
背景/目的：皮肤鳞状细胞癌（cSCC）是头颈部常见的皮肤恶性肿瘤，易向局部淋巴结转移。上皮-间质转化（EMT）和癌症相关成纤维细胞（CAFs）在疾病进展中具有明确作用。本研究构建并表征了由患者来源的转移性cSCC细胞系（各具不同表型）与真皮或淋巴结来源的成纤维细胞组成的多细胞肿瘤球体（MCTS），旨在探究不同细胞组合如何影响MCTS的形成、空间结构及侵袭行为。我们假设不同cSCC与成纤维细胞的相互作用会对球体形成和侵袭产生差异化影响。 方法：通过活细胞显微镜技术评估各细胞系-成纤维细胞组合的特异性空间结构，检测EMT及CAF标志物的表达。进一步利用MCTS侵袭模型，探究侵袭模式与癌细胞系EMT表型之间的关联。 结果：研究发现转移性cSCC/成纤维细胞MCTS可自组织形成独特的空间结构。其在胶原中的侵袭行为虽受成纤维细胞影响，但主要取决于来源癌细胞的EMT状态。 结论：这些发现凸显了MCTS作为研究转移性cSCC中肿瘤-基质相互作用及侵袭动力学的模型具有生理相关性和实用价值。
肿瘤(癌症)患者之家