To explore the molecular mechanism of microRNA-599(miR-599) in the migration and invasion of glioma. Clinicopathological characteristics of 33 patients were analyzed. Based on reverse transcription-PCR, miR-599 was down-regulated in glioma tissues compared with adjacent normal brain tissues (P < 0.001). Moreover, negative correlations between miR-599 and periostin protein expression in glioma tissues (P < 0.01) and necrosis by magnetic resonance imaging (P < 0.05) were observed. Transwell and wound healing assays showed that overexpression of miR-599 inhibited glioma cell migration and invasion. miR-599 down-regulated periostin expression by targeting the 3'-untranslated region. Additionally, re-expression of periostin partial reversed the suppressive effect of miR-599 on migration and invasion in vitro and in vivo. microRNA-599 inhibits proliferation and invasion by down-regulating periostin expression in vitro and in vivo.