Objective: To study the role of microRNA-301a (miR-301a) in glioma. Methods: Real-time quantitative RT-PCR (qRT-PCR) was carried out to detect the miR-301a expression in tissues. CCK-8 assay and cell viability assay were used to determine cell proliferation and temozolomide (TMZ) resistance. The association between mir-301a and BTG1 was confirmed by luciferase reporter assay as well as Western blot. Results: The miR-301a expression was raised in glioma significantly. Obvious interrelation was detected between high miR-301a expression level with high WHO grade. The 5-year overall survival (OS) rate was dramatically reduced in patients with high miR-301a expression. Besides, miR-301a was a prognostic variable for OS rate. Moreover, miR-301a overexpression promoted glioblastoma cell proliferation and TMZ resistance through regulating BTG1. Conclusions: High miR-301a expression in glioma is participated in glioblastoma cell proliferation as well as TMZ resistance, which may help for the prognosis and treatment for glioma.