Shimpi Adrian A, Williams Erik D, Ling Lu, Tamir Tigist, White Forest M, Fischbach Claudia
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853, United States.
Department of Information Science, Cornell University, Ithaca, New York 14853, United States.
ACS Biomater Sci Eng. 2023 Dec 11;9(12):6835-6848. doi: 10.1021/acsbiomaterials.3c01034. Epub 2023 Nov 28.
Increased fibrotic extracellular matrix (ECM) deposition promotes tumor invasion, which is the first step of the metastatic cascade. Yet, the underlying mechanisms are poorly understood as conventional studies of tumor cell migration are often performed in 2D cultures lacking the compositional and structural complexity of native ECM. Moreover, these studies frequently focus on select candidate pathways potentially overlooking other relevant changes in cell signaling. Here, we combine a cell-derived matrix (CDM) model with phosphotyrosine phosphoproteomic analysis to investigate tumor cell migration on fibrotic ECM relative to standard tissue culture plastic (TCP). Our results suggest that tumor cells cultured on CDMs migrate faster and in a more directional manner than their counterparts on TCP. These changes in migration correlate with decreased cell spreading and increased cell elongation. While the formation of phosphorylated focal adhesion kinase (pFAK)+ adhesion complexes did not vary between TCP and CDMs, time-dependent phosphoproteomic analysis identified that the SRC family kinase LYN may be differentially regulated. Pharmacological inhibition of LYN decreased tumor cell migration and cytoskeletal rearrangement on CDMs and also on TCP, suggesting that LYN regulates tumor cell migration on CDMs in combination with other mechanisms. These data highlight how the combination of physicochemically complex in vitro systems with phosphoproteomics can help identify signaling mechanisms by which the fibrotic ECM regulates tumor cell migration.
增加的纤维化细胞外基质（ECM）沉积促进肿瘤侵袭，这是转移级联反应的第一步。然而，由于传统的肿瘤细胞迁移研究通常在缺乏天然ECM组成和结构复杂性的二维培养中进行，其潜在机制仍知之甚少。此外，这些研究经常聚焦于特定的候选途径，可能忽略了细胞信号传导中的其他相关变化。在这里，我们将细胞衍生基质（CDM）模型与磷酸酪氨酸磷酸化蛋白质组分析相结合，以研究肿瘤细胞在纤维化ECM上相对于标准组织培养塑料（TCP）的迁移情况。我们的结果表明，在CDM上培养的肿瘤细胞比在TCP上培养的肿瘤细胞迁移得更快且更具方向性。这些迁移变化与细胞铺展减少和细胞伸长增加相关。虽然磷酸化粘着斑激酶（pFAK）+粘着复合物的形成在TCP和CDM之间没有差异，但时间依赖性磷酸化蛋白质组分析表明，SRC家族激酶LYN可能受到不同的调节。LYN的药理学抑制降低了肿瘤细胞在CDM上以及在TCP上的迁移和细胞骨架重排，表明LYN与其他机制共同调节肿瘤细胞在CDM上的迁移。这些数据突出了物理化学复杂的体外系统与磷酸化蛋白质组学的结合如何有助于识别纤维化ECM调节肿瘤细胞迁移的信号传导机制。
