Study on the molecular mechanism of chlorotoxin from Buthus martensii Karsch in the inhibition of glioma angiogenesis

BACKGROUND: Glioma is a highly malignant intracranial tumor with dismal prognosis. Tumor angiogenesis critically drives glioma growth and invasion, making vascular endothelial cells a promising therapeutic target. Buthus martensii Karsch chlorotoxin (BmK CT) presents favorable antitumor properties, whereas its regulatory role in glioma angiogenesis and the specific mechanism remain poorly defined. AIM: To investigate the regulatory role of BmK CT in glioma angiogenesis and the underlying mechanism. METHODS: HUVECs were cultured alone or exposed to glioma conditioned medium prepared from primary glioma cells. Functional assays were performed to assess the impacts of BmK CT on endothelial proliferation, migration and tube formation. Quantitative proteomics and bioinformatics analysis were applied to screen differentially expressed proteins (DEPs) and explore underlying mechanisms. RESULTS: BmK CT significantly inhibited the proliferation of HUVEC cells and reduced their migration and tube formation abilities. BmK CT reversed the promoting effect of primary glioma cells on the migration of HUVEC cells. Proteomics identified multiple DEPs. Enrichment analysis suggested that BmK CT exerts an anti-glioma angiogenesis effect through a multi-target mechanism by inhibiting pro-angiogenic pathways, blocking energy metabolism, and regulating the inflammatory microenvironment. Furthermore, expression pattern clustering analysis revealed six DEPs expression patterns. Parallel reaction monitoring (PRM) analysis supported nine candidate proteins (THBS1, THBS4, COMP, F2, A2M, C3, RIGI, EZR, and ITGA6) associated with glioma angiogenesis. Conclusion: This study demonstrated in vitro that BmK CT inhibits endothelial angiogenic behaviors relevant to glioma progression and modulates the tumor microenvironment, and identified nine candidate DEPs, providing a theoretical basis for future molecular targeted therapy strategies for glioma.