MicroRNAs, small non-coding RNAs (21-23 nt in length), are known for regulating carcinogenesis and tumor progression in glioma. MicroRNA-874 (miR-874) has shown play an important role in many human cancers as tumor suppressors. Our previous studies found that miR-874 reduced in glioma tissue and overexpression of miR-874 inhibits glioma cell proliferation and induces apoptosis. However, the biological functions of miR-874 in chemotherapy sensitivity enhancing effect have not been elucidated completely. The present study evaluated the biological function and mechanism of miR-874 in chemotherapy sensitivity enhancing effect in glioma. Our results revealed that the cell proliferation and migration were decreased, cell apoptosis was induced, and the mitochondrial membrane potential was also declined in glioma cells treated with miR-874 with temozolomide (TMZ). And miR-874 with TMZ can reduce the expression level of migration related protein MMP-2 and MMP-9, down-regulate the expression level of Bcl-2 and induce the expression level of Bax. More importantly, miR-874 with TMZ reduced the expression level of STAT3 protein and inhibited STAT3 phosphorylation. Our results demonstrated that overexpression of miR-874 potentiates chemosensitivity of glioblastoma to TMZ by the oncogenic STAT3 pathway, which might provide novel strategies for clinical treatment.