<b>Abstract</b>— <p>Melanoma is one of the most common cancers in the world, especially in young adults. Due to the emergence of the resistance for treatments, the identification of novel potential molecular targets is of specific importance. Previously, we have identified functional mechanosensitive Piezo1 channels in the plasma membrane of aggressive melanoma SK-MEL-2 cell line and evidenced that selective Piezo1 activation by chemical agonist Yoda1 completely prevented melanoma spheroid formation. Here, we expanded the number of experimental tests to reveal the role of Piezo1 activity in pathophysiological reactions of SK-MEL-2 cells that are linked with cell motility and migration. In particular, we assessed the melanoma cell motility using wound closure and spheroid reactivation assay in the presence of Piezo1-selective chemical activator, Yoda1. Ca²⁺ imaging allowed us to detect variations in a number of functionally active Piezo1 between the cells on the surface of melanoma spheroids. The processes of melanoma spheroid reactivation were unaffected by Yoda1. At the same time, wound healing assay showed a decrease of SK-MEL-2 migratory activity in the presence of Yoda1. Our results demonstrate differential roles of Piezo1 in regulation of migratory properties of melanoma cells.</p>