Background: Glioblastoma (GBM) is an aggressive brain cancer with limited treatment options and a poor prognosis. Voltage-gated sodium channels, particularly Nav1.6 (SCN8A), have been implicated in tumor progression across various cancers. This study investigates the role of Nav1.6 in promoting GBM proliferation and migration via Na⁺/H⁺ exchanger-1 (NHE1), and its regulation of ERK and AKT signaling pathways to identify potential therapeutic strategies.
Methods and results: Immunohistochemistry revealed high Nav1.6 expression in glioma tissues, peaking in grade IV GBM. Human GBM cell lines (U251, U138, U87) were used to explore the functional roles of Nav1.6 and NHE1 through siRNA-mediated silencing and pharmacological modulation. Quantitative PCR and Western blot confirmed the effective silencing of both targets and the downstream effects on protein expression. CCK8 and EdU DNA cell proliferation assays showed that inhibition of Nav1.6 or NHE1 significantly suppressed cell proliferation. Wound healing and Transwell migration assays demonstrated reduced cell motility upon gene silencing or treatment with TTX and EIPA. Flow cytometry and Caspase-3 analysis confirmed enhanced apoptosis following dual inhibition. Conversely, activation of Nav1.6 by ATXII promoted proliferation and migration, effects reversed by NHE1 downregulation. Importantly, combined suppression of Nav1.6 and NHE1 led to significant downregulation of phosphorylated AKT and ERK1/2, suggesting their cooperative role in modulating survival signaling in GBM.
Conclusion: Nav1.6 promotes GBM progression by regulating ion homeostasis via NHE1 and activating AKT/ERK pathways. Dual targeting of Nav1.6 and NHE1 may offer a promising strategy to inhibit proliferation and induce apoptosis in GBM.
Keywords: ERK and AKT pathway; Glioblastoma; Na+/H+ exchanger1 (NHE1); NaV1.6; Voltage-gated sodium channels.
© 2025. The Author(s), under exclusive licence to Springer Nature B.V.