Head and neck squamous cell carcinoma (HNSCC) is a prevalent cancer type, with cisplatin being a primary treatment approach. However, drug resistance and therapy failure pose a significant challenge, affecting nearly 50% of patients over time. This research had two aims: (1) to optimize a 3D cell-culture method for assessing the interplay between tumor cells and cancer-associated fibroblasts (CAFs) in vitro; and (2) to study how cisplatin impacts the Notch pathway, particularly considering the role of CAFs. Using our optimized “3D sheet model” approach, we tested two HNSCC cell lines with different cisplatin sensitivities and moderate, non-mutated NOTCH1 and -3 expressions. Combining cisplatin with a γ-secretase inhibitor (crenigacestat) increased sensitivity and induced cell death in the less sensitive cell line, while cisplatin alone was more effective in the moderately sensitive line and sensitivity decreased with the Notch inhibitor. Cisplatin boosted the expression of core Notch signaling proteins in 3D monocultures of both lines, which was counteracted by crenigacestat. In contrast, the presence of patient-derived CAFs mitigated effects and protected both cell lines from cisplatin toxicity. Elevated NOTCH1 and NOTCH3 protein levels were consistently correlated with reduced cisplatin sensitivity and increased cell survival. Additionally, the Notch ligand JAG2 had additional, protective effects reducing cell death from cisplatin exposure. In summary, we observed an inverse relationship between NOTCH1 and NOTCH3 levels and cisplatin responsiveness, overall protective effects by CAFs, and a potential link between JAG2 expression with tumor cell survival.
头颈部鳞状细胞癌（HNSCC）是一种常见的癌症类型，顺铂是其主要的治疗手段。然而，耐药性和治疗失败构成了重大挑战，长期来看影响了近50%的患者。本研究有两个目标：（1）优化一种三维细胞培养方法，用于在体外评估肿瘤细胞与癌症相关成纤维细胞（CAFs）之间的相互作用；（2）研究顺铂如何影响Notch通路，特别是考虑到CAFs的作用。通过我们优化的“三维片层模型”方法，我们测试了两种具有不同顺铂敏感性的HNSCC细胞系，这些细胞系具有中等且未突变的NOTCH1和NOTCH3表达。将顺铂与γ-分泌酶抑制剂（crenigacestat）联合使用，提高了低敏感性细胞系的敏感性并诱导细胞死亡，而顺铂单独使用在中等敏感性细胞系中更为有效，且Notch抑制剂降低了其敏感性。顺铂增强了两种细胞系在三维单培养中核心Notch信号蛋白的表达，而crenigacestat则抵消了这一效应。相比之下，患者来源的CAFs的存在减轻了这些效应，并保护两种细胞系免受顺铂毒性。NOTCH1和NOTCH3蛋白水平的升高与顺铂敏感性降低和细胞存活率增加持续相关。此外，Notch配体JAG2具有额外的保护作用，减少了顺铂暴露引起的细胞死亡。总之，我们观察到NOTCH1和NOTCH3水平与顺铂反应性呈负相关，CAFs具有总体保护作用，并且JAG2表达与肿瘤细胞存活之间存在潜在联系。
肿瘤(癌症)患者之家