Cancer Cells Traverse Faster in Confined Space by Modifying Vimentin filaments With Nuclear Deformation and Promoting the Growth of Desired Tumor Spheroids

Migrating cancer cells restructure their cytoskeleton to form protrusive structures, aiding diverse migration modes through confined spaces. These migration modes enhance cancer cells' capacity to navigate the constrained barriers of the metastatic cascade and facilitate distant migration toward preferred metastatic destinations. Vimentin's role in this process and its impact on post-migration tumor spheroid growth remain understudied. Using a novel microfluidic setup, this study explored how cell and nuclear stiffness affect cancer cell migration in confined spaces. Results showed cells alter Vimentin intermediate (VIM) filaments, deform nuclei, and use bleb-based protrusions for faster movement. A trans-well system confirmed these findings and revealed reduced tumor spheroid growth rates in cells migrating through confined spaces. While maintaining cell and nuclear stiffness gene expression, post-migration spheroids exhibited varied Extracellular matrix (ECM), integrin, and stemness gene profiles. Collectively, this study showed that lung cancer cells migrating swiftly in confined spaces maintained stiffness-related genes but regulated ECM, integrin, and stemness genes differently to form tumor spheroids. © 2025 The Author(s).