MicroRNAs (miRNAs) regulate biogenesis and disease development by targeting numerous mRNAs. miRNA (miR)-124 and its direct target, inhibitor of apoptosis-stimulating protein of p53 (iASPP), may be involved in tumor development and progression. The aim of the present study was to explore the role of miR-124-targeted iASPP in glioma. The results demonstrated that miR-124 was aberrantly expressed in astrocytic glioma tissue and in the human glioblastoma cell lines U87 and U251. The expression of miR-124 was lower in astrocytic gliomas compared with normal brain (NB) tissues, with a more reduced expression in higher-grade tumors. In addition, several miR-124 loci (including miR-124-1, miR-124-2 and miR-124-3) were revealed to be more highly methylated in U87 cells compared with methylation levels in U251 cells and NB cells. Furthermore, the expression of iASPP was higher in high-grade astrocytic gliomas compared with low-grade astrocytic gliomas. miR-124 overexpression effectively inhibited U87 and U251 cell migration. In addition, miR-124 regulated cell viability and arrested the cell cycle at the G0/G1 phase in these two cell lines. miR-124 also reduced the expression levels of the cell cycle related genes iASPP, cyclin-dependent kinase (CDK)4, CDK6 and cyclin D1. Results from the present study indicated that expression of the miR-124 target gene iASPP may contribute to glioma development and progression.