Objective Voltage-gated sodium channels (VGSCs) are expressed in glioma U251 cells and affect the proliferation, invasion and apoptosis of U251 cells. It is reasonable to hypothesize that the cysteine-rich buccal gland protein (CRBGP), a VGSCs blocker isolated from the buccal glands of Lampetra japonica, may suppress the activity of U251 cells. Methods Firstly, the proliferation of U251 cells in the presence of CRBGP was detected by MTT assay. And the morphology, cytoskeleton and nucleus of U251 cells after treated with CRBGP were observed by Wright-Giemsa, FITC-phalloidin and Hoechst 33258 staining assays, respectively. Subsequently, extracellular matrix proteins such as collagen IV, fibronectin and laminin were used to detect the effect of CRBGP on U251 cells’ adhesion. In addition, the migration and invasion of U251 cells treated with CRBGP were detected by transwell assays. And the internal mechanisms of CRBGP on U251 cells’ apoptosis and mobility were explored by Western blot. Finally, the anti-inflammatory effect of CRBGP on U251 cells was detected by enzyme linked immunosorbent assay. Results CRBGP inhibited the proliferation of U251 cells by inducing apoptosis in a mitochondrial-dependent pathway and preventing the release of proinflammatory factor interleukin-6. Also, the VGSCs blocking and anti-inflammatory activities of CRBGP contributed to its inhibitory effects on the adhesion, migration and invasion of U251 cells. Finally, CRBGP affected the proliferation and mobility of U251 cells through the suppression of the FAK-AKT pathway. Conclusion Together, our data indicated that CRBGP could exhibit its anti-tumor activity probably by its VGSCs inhibitory property, providing a basis for the functional information of VGSCs to the gliomas.