Glioma is the most common malignant tumor of the adult central nervous system, with high malignancy and mortality. The combination of temozolomide (TMZ) chemotherapy and radiotherapy is currently the standard first-line regimen for high-grade gliomas, but with the progress of chemotherapy, most patients will eventually develop resistance to TMZ and lead to glioma treatment failure. The results of this study provide TMZ-resistant glioma cell lines for basic and clinical research and provide potential molecular targets for reversing TMZ resistance and enhancing the effectiveness of TMZ therapy, which are of great significance for the treatment of gliomas and the improvement of patient prognosis.