Increased migratory, invasive and metastatic potential is one of the main pathophysiological determinants of malignant cells. Mechanosensitive calcium-permeable ion channels are among the key membrane proteins that participate in processes of, cellular motility. Local calcium influx via mechanosensitive channels was proposed to regulate calcium-dependent molecules involved in cell migration. Piezo transmembrane proteins were shown to act as calcium-permeable mechanosensitive ion channels in various cells and tissues, including a number of tumor cells. Furthermore, an elevated expression of Piezo1 is correlated with poor prognosis for some types of cancers. At the same time, functional impact of Piezo1 channels on pathophysiological reactions of tumor cells remains largely unknown. Here, we used 3T3B-SV40 mouse fibroblasts as a model to study the effect of Yoda1, selective Piezo1 activator, on migrative properties of transformed cells. RT-PCR and immunofluorescent staining showed the presence of native Piezo1 in 3T3B-SV40 fibroblasts. Functional expression of Piezo1 in plasma membrane of 3T3B-SV40 cells was confirmed by calcium measurements and single channel patch-clamp analysis. Particularly, application of Yoda1 resulted in rapid calcium influx and induced typical channel activity in membrane patches with characteristics identical to stretch-activated channels in 3T3B-SV40 cells. Importantly, dose-dependent inhibition of cellular migration by Yoda1 was found in wound healing assay using live cell imaging. Consistently, microscopic analysis showed that Yoda1 significantly altered cellular morphology, induced F-actin assembly and stress fiber formation indicating partial reversion of transformed phenotype. The results demonstrate for the first time that Piezo1 activation by selective agonist Yoda1 could be favorable for inhibiting migrative potential of transformed cells with native Piezo1 expression. (C) 2019 Elsevier Inc. All rights reserved.