Electrical stimulation induces differentiation onset consistent with a therapeutic approach in neuroblastoma cells.

Cancer therapies based on differentiation processes are strategies that try to induce the maturation of cancer cells into a differentiated and not proliferating state, aiming in this way to arrest the tumor growth. While conventional therapies often lack specificity and can harm normal cells, differentiation strategies have shown to be promising but they still remain limited in clinical application. Electrical stimulation (ES), which should not be confused with electroporation, has been shown to induce specific cellular responses, including differentiation. Here we showed that voltage-controlled biphasic pulses at 500 mV/mm and 100 Hz suppresses proliferation and promotes neuronal differentiation in the neuroblastoma cell line N2a, both in monolayer and 3D neurosphere cultures. The morphological changes induced by ES associated with neuronal differentiation, downregulated proliferation markers (H3S10ph and Ki-67), modulated neuronal differentiation markers (Neurod1 and SOX2) and reduced colony and neurosphere formation, all this without causing DNA damage. Therefore, ES represents a non-genotoxic mechanism to halt tumor cell growth, distinguishing it from standard cytotoxic therapies. These findings suggest that ES offers a means to suppress proliferation and induce differentiation in neuroblastoma cells, providing a potential foundation for less harmful therapeutic strategies targeting this pediatric cancer.