In this study, we investigated the role of Piezo1 channel in regulating the [Ca2+]i, cofilin phosphorylation and filopodial formation and thereby mediating matrix stiffness regulation of lung cancer cell migration in A549 cells, a NSCLC line, growing on soft and stiff polyacrylamide (PA) hydrogels with stiffnesses close to these of healthy and cancerous lung tissues, respectively, and aim to gain mechanistic understanding of how the Piezo1 channel is involved in the regulation of lung cancer cell migration by ECM. This evidence concerns the gene CFL1 and lung cancer.