Mechanical interactions of invasive cancer cells through their substrate evolve from additive to synergistic.

Non-contacting, adjacent cancer cells can mechanically interact through their substrate to increase their invasive and migratory capacities that underly metastases-formation. Such mechanical interactions may induce additive or synergistic enhancement of invasiveness, potentially indicating different underlying force-mechanisms. To identify cell-cell-gel interactions, we monitor the time-evolution of three-dimensional traction strains induced by MDA-MB-231 breast cancer cells adhering on physiological-stiffness (1.8 kPa) collagen gels and compare to simulations. Single metastatic cells apply strain energies of 0.2-2 pJ (average 0.51 ± 0.06 pJ) at all observation times (30-174 min) inducing a mechanical volume-of-effect in the collagen gel that is initially (<60 min from seeding) on the cell-volume scale (∼3000 µm) and on average increases with time from cell seeding. When cells adhere closely adjacent, at short times (<60 min) we distinguish the additive contributions of neighboring cells to the strains, while at longer times strain fields are synergistically amplified and may facilitate increased cooperative/collective cancer-cell-invasiveness. The results of well-spaced and closely adjacent cells at short times match our simulations of additive deformations induced by radially applied strains with experimentally based inverse-distance decay. We thus reveal a time-dependent evolution from additive to synergistic interactions of adjacently adhering cells that may facilitate metastatic invasion.