Gliomas are characterized by high mortality and disability rates. Cancer-testis antigens (CTAs) are among the most promising therapeutic targets for combating cancer. While several CTAs have been associated with the development and progression of gliomas, the role of ATPase family AAA domain-containing protein 2 (ATAD2) in this context has not been thoroughly investigated. In this study, both in vitro and in vivo experiments validated the role of ATAD2 in enhancing malignant phenotypes. The LN229 cell lines were employed for RNA-seq and proteomics to uncover downstream targets of ATAD2. Results showed that elevated ATAD2 expression was noted in glioblastoma (GBM). ATAD2 knockdown significantly reduced the proliferation, migration, and invasion capabilities of GBM cells, while its overexpression had the opposite effect. The knockdown of ATAD2 led to a decrease in subcutaneous tumor size and weight, a reduction in Ki67 expression, and an extension of survival in mice bearing intracranial in situ tumors. Mechanistically, a positive feedback loop involving ATAD2 and E2F transcription factor 1 (E2F1) was identified to enhance the transcriptional activation of pyruvate dehydrogenase kinase 1 (PDK1). Notably, the expression levels of these genes were found to be positively correlated, with patients exhibiting high levels of these genes tending to have poorer prognoses. These findings demonstrate that ATAD2 plays a pivotal role in the malignant progression of glioma and synergizes with E2F1 to promote PDK1 expression, suggesting its potential as a therapeutic target for glioma.