This study finds that knockdown of CLC-3 inhibits p65 nuclear translocation, decreases transcriptional activity of NF-κB, and reduced MMP-3/9 expression, indicating that during metastasis, glioma cells not only passively regulate cellular volume and morphology to navigate the constrained space via CLC-3 chloride channel-mediated Cl− efflux, but also actively create an extracellular microenviroment that favors invasion through CLC-3 channels. Here, CLCN3 is linked to central nervous system cancer.