The present study was designed to investigate the function of matrix metalloproteinase‑9 (MMP‑9) in human glioma cells and the potential regulatory mechanisms. Reverse transcription‑quantitative polymerase chain reaction was used to analyze the expression of MMP‑9 and microRNA‑34a (miR‑34a) in the plasma of patients with glioma and healthy volunteers. MTT and Transwell assays were used to assess cell growth and migration, respectively. Annexin‑V/propidium iodide staining was used to measure cell apoptosis. In addition, MMP‑9 expression was measured using western blot analysis. In patients with glioma, MMP‑9 expression was increased, while miR‑34a expression was suppressed, compared with the normal group. Overall survival (OS) and disease‑free survival (DFS) of patients with high MMP‑9 expression were decreased compared with those with low MMP‑9 expression. OS and DFS of patients with low miR‑34a expression were decreased compared with those with high miR‑34a expression. Downregulation of miR‑34a promoted cell growth and migration, and inhibited apoptosis in U251‑MG glioma cells. However, overexpression of miR‑34a inhibited cell growth and migration, and induced apoptosis in glioma cells. Furthermore, downregulation of miR‑34a using anti‑miR‑34a induced MMP‑9 protein expression in glioma cells; whereas, overexpression of miR‑34a suppressed MMP‑9 protein expression in glioma cells. SB‑3CT, an inhibitor of MMP‑9, attenuated the effects of miR‑34a mimic on glioma cells. Together, these results indicated that miR‑34a inhibited cell growth and migration in human glioma cells by regulating MMP‑9.