虽然增强的肿瘤细胞迁移是肿瘤传播的关键过程，但对肿瘤细胞与机械限制之间因果关系的机制了解仍然有限。在这里，我们将微流体平台的使用与基因组工具结合起来，以表征受限细胞迁移，系统地揭示与乳腺癌 (BC) 细胞迁移表型相关的全局信号图谱。在 3D 微流体中评估了 7 种 BC 细胞系的自发迁移能力设备及其迁移能力与公开可用的分子特征相关。已确定的信号通路对调节 BC 迁移能力的作用是通过配体结合刺激受体或通过 siRNA 沉默抑制来确定的。在微流体装置中评估下游对细胞迁移的影响，同时通过 RT-qPCR 监测分子变化。715 个基因的表达与 BC 细胞迁移表型相关，表明 TNF-α 是最重要的上游调节因子之一。信号转导实验表明，TNF-α 刺激三阴性、间质样 BC 细胞的有限迁移，这些细胞也具有高 TNFR1 表达的特征，但抑制具有低 TNFR1 表达的上皮样细胞的迁移。 TNFR1 与迁移能力和三阴性间充质表型密切相关。 TNF/TNFR1 信号传导的下游，NFKB 的转录调控似乎对于驱动细胞在有限空间内迁移很重要。TNF-α/TNFR1 信号轴在驱动 BC 细胞有限迁移中发挥着关键作用，成为一种有前途的靶向治疗策略三阴性间充质 BC 细胞的传播和转移。版权所有 © 2024。由 Elsevier Inc. 出版。

While enhanced tumor cell migration is a key process in the tumor dissemination, mechanistic insights into causal relationships between tumor cells and mechanical confinement are still limited. Here we combine the use of microfluidic platforms to characterize confined cell migration with genomic tools to systematically unravel the global signaling landscape associated with the migratory phenotype of breast cancer (BC) cells.The spontaneous migration capacity of seven BC cell lines was evaluated in 3D microfluidic devices and their migration capacity was correlated with publicly available molecular signatures. The role of identified signaling pathways on regulating BC migration capacity was determined by receptor stimulation through ligand binding or inhibition through siRNA silencing. Downstream effects on cell migration were evaluated in microfluidic devices, while the molecular changes were monitored by RT-qPCR.Expression of 715 genes was correlated with BC cells migratory phenotype, revealing TNF-α as one of the top upstream regulators. Signal transduction experiments revealed that TNF-α stimulates the confined migration of triple negative, mesenchymal-like BC cells that are also characterized by high TNFR1 expression, but inhibits the migration of epithelial-like cells with low TNFR1 expression. TNFR1 was strongly associated with the migration capacity and triple-negative, mesenchymal phenotype. Downstream of TNF/TNFR1 signaling, transcriptional regulation of NFKB seems to be important in driving cell migration in confined spaces.TNF-α/TNFR1 signaling axis reveals as a key player in driving BC cells confined migration, emerging as a promising therapeutic strategy in targeting dissemination and metastasis of triple negative, mesenchymal BC cells.Copyright © 2024. Published by Elsevier Inc.