• Expression of Cathepsin L and CD133 were correlated with human glioma malignancy.• Knockdown of Cathepsin L leads to radiosensitization in GSCs.• The improved radiosensitivity was through stemness, DNA repair, and apoptosis.• Knockdown of Cathepsin L exhibited less tumor growth after radiation in vivo.• Cathepsin L is a promising therapeutic target for clinical therapy in GBM patients.
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