Effect of Piezo1 activation and CYRI-B loss in melanoma cell membrane dynamics

Background Cancer cell migration is a critical step in metastasis, driven by cytoskeletal remodelling in response to extracellular mechanical cues. Piezo1, a mechanosensitive ion channel, has been known to promote cell motility through calcium-dependent activation of actin-regulatory pathways. In parallel, CYRI-B acts as a local inhibitor of lamellipodial protrusion by binding Rac1 to prevent activation of the Scar/WAVE complex. However, whether these two pathways interact to regulate cancer cell motility remains unexplored. Methods Using B16F1 mouse melanoma cells, we investigated how Piezo1 activation (via the small molecule agonist Yoda1) and CYRI-B knockout influence collective cell migration and membrane fluctuations during cell spreading. Scratch-wound healing assays quantified collective migration, and phase-contract live-cell imaging with kymograph analysis assessed spreading and protrusion behaviour in both wild-type and CYRI-B knockout cells with and without treatment with Yoda1. Results Piezo1 activation significantly increased collective cell migration by approximately 35%, cell area by approximately 30%, and membrane fluctuations by approximately two-fold in wild-type cells. However, none of these changes were observed after Piezo1 activation in CYRI-B knockout cells. Kymograph analysis showed that protrusion length increased along with protrusion angle after Piezo1 activation in the presence of CYRI-B. However, these increases were also not observed in CYRI-B knockout cells. Conclusion Our findings show that CYRI-B may influence the Piezo1-mediated increase in cancer cell migration and membrane dynamics. This raises the possibility of a previously unrecognized connection between mechanosensitive Piezo1 activation and CYRI-B-mediated membrane fluctuations, adding a new dimension to membrane dynamics in cancer cell motility and spreading.