Objective · To explore the role and underlying mechanisms of mitochondrial dicarboxylate carrier SLC25A10 in the occurrence and progression of glioma. Methods · The U251 glioma cell lines over-expressing or depleted of SLC25A10 were obtained via lentivirus infection. Western blotting and real-time PCR were used to measure the efficiency of knockdown and over expression of SLC25A10. The effects of SLC25A10 on proliferation and tumorigenicity of U251 cells were detected with cell counting, CCK-8 assay, clonogenic assay, and tumor formation in nude mice. The ATP level and the activity of AMPK and its downstream mTOR signaling pathway were detected with kits and Western blotting, respectively. Results · Knock down of SLC25A10 decreased the proliferation, clone number, tumorigenicity, and ATP level in U251 cells. However, over expression of SLC25A10 didn’t significantly affect the proliferation of U251 cells, but increased the ATP level and the radiation resistance. Study showed that knock down of SLC25A10 activated AMPK, which in turn inhibited mTOR signaling pathway, resulting in decreased cell proliferation and increased cell sensitivity to radiotherapy. Conclusion · Knock down of SLC25A10 inhibits the mTOR signaling pathway, slows down glioma proliferation, and increases cell sensitivity to radiotherapy. These findings provide a potential target for the treatment of glioma.