In order to further study the mechanism of chemotherapy resistance in human gliomas and explore potential therapeutic methods, we investigated the effects of tumor necrosis factor α-induced protein 1 (TNFAIP1) on migration and invasion. Reverse transcription PCR and Western blot assays showed that temozolomide increased the expression of TNFAIP1. We overexpressed TNFAIP1 in U251 cells, then we found it could promote the inhibition of temozolomide (TMZ) on migration and invasion of U251 cells, and this effect was achieved by inhibiting epithelial-to-mesenchymal transition (EMT) signaling pathway. This finding is helpful to provide a new theoretical basis for clinical treatment of human glioma and provide a new strategy for its treatment.