Endogenously expressed Piezo1 activation enhances migration in glioblastoma cells

Glioblastoma is a highly invasive brain tumor in which microenvironmental cues, including extracellular matrix composition and mechanical stimuli, play a critical role in regulating cell migration. Mechanosensitive ion channels such as Piezo1 have emerged as key mediators of cellular responses to mechanical signals; however, their functional contribution to glioblastoma motility remains incompletely understood. In this study, we investigated the role of endogenously expressed Piezo1 in glioblastoma cells using the U-251 MG cell line with a combination of wound healing assays for cell migration tracking, electrophysiology, Ca²⁺ imaging, pharmacological modulation, and gene expression analysis. Laminin-coated substrates significantly enhanced cell migration, indicating that extracellular matrix composition influences glioblastoma motility. Quantitative PCR analysis revealed that Piezo1, but not Piezo2, is expressed in U-251 MG cells. Functional activity of Piezo1 was confirmed by application of the agonist Yoda1, which induced robust inward currents and intracellular Ca²⁺ signals. Pharmacological profiling demonstrated that the Yoda1 analogue Dooku1 selectively attenuated Yoda1-induced Piezo1 activation, whereas the alternative activator Jedi1 and the mechanosensitive channel inhibitor GsMTx4 showed no significant effects under our experimental conditions, highlighting cell-type-specific differences in Piezo1 modulation. Prolonged Yoda1 stimulation reduced Piezo1 mRNA expression, suggesting a potential feedback mechanism. Importantly, activation of Piezo1 significantly enhanced glioblastoma cell migration, while co-application of Dooku1 abolished this effect. In contrast, Dooku1 alone did not affect migration. Together, these findings demonstrate that Piezo1 is functionally expressed in glioblastoma cells and that its activation promotes migratory behavior. Our results identify mechanosensitive Piezo1 signaling as a potential contributor to glioblastoma progression and suggest that it may represent a context-dependent pharmacological target.