Mechano-stress endorsing heterogeneous lung cancer cells migration into confined channels and investigating tumor spheroids growth of confined space migrating cells

Cancer cells in the tumor microenvironment and in the metastatic cascade leverage their different intrinsic and extrinsic properties to overcome the confined barriers of the metastatic cascade during metastasis. Although various studies have focused on cell migration under confinement, how the heterogeneity in the stiffness levels of single lung cancer cells affects their migration into confined spaces and the growth of tumor spheroids of confined space migrating cells remains unknown. This study explores how cancer cells with varying stiffness levels steer confined spaces and form post-migration tumor spheroids. Using a single-cell migration platform and a confined trans-well migration setup, the research uncovers that lung cancer cells with lower stiffness and reduced VIM expression exhibit selective migration into confined spaces. These cells, forming irregularly shaped spheroids post-migration, display nuclear shape deformation and downregulated VIM and LMNA genes linked to cell and nuclear stiffness. The findings highlight the heterogeneity of cancer cell migration in confined environments and the subsequent growth of tumor spheroids. © The Author(s) 2026.