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Background/Aims: The transcription factor Krüppel-like factor (KLF) 8 plays important roles in tumorigenesis and tumor metastasis. However, the relationship between KLF8 and glioma cell chemoresistance is not known. Methods: The effects of KLF8 on glioma cell proliferation, apoptosis and chemosensitivity to temozolomide (TMZ) were analyzed by Cell Counting Kit 8 assay and flow cytometry assay. A xenograft model was used to study the effect of KLF8 on tumor growth and sensitivity to TMZ. Results: We found that in the absence of KLF8, glioma cells showed greater sensitivity to TMZ, resulting in the inhibition of cell growth and enhanced apoptosis. KLF8 overexpression had the opposite effect; that is, cell resistance to TMZ was increased, which was associated with β-catenin activation. Conclusion: Taken together, these data suggest that KLF8 modulates glioma cell resistance to TMZ via activation of β-catenin; therefore, therapies that inhibit KLF8 levels in glioma can enhance the efficacy of TMZ treatment.