Invasion of tumor cells is the primary cause of therapeutic failure in the treatment of malignant gliomas. In an attempt to investigate the properties of the malignant progression of glioma cells, we examined the correlation between cell migration and glial cell-derived neurotrophic factor (GDNF) secretion of two glioma cell lines which differ in their invasive phenotypes. Here, we show that the high-grade C6 cells are more migrative and secrete more GDNF than the low-grade Hs683 cells. GDNF signaling is more highly activated in C6 cells than in Hs683 cells. Treatment of the Hs683 cells with GDNF significantly increased migration comparable to the C6 cells, revealing the autocrine and/or paracrine effect of GDNF on promotion of the glioma cell migration. We then examined the involvement of mitogen-activated protein kinases (MAPKs) including c-Jun N-terminal protein kinase (JNK), extracellular signal-regulated kinases (ERKs) and p38 MAPK in Hs683 cell migration induced by GDNF. A prominent activation of JNK, ERKs and p38 MAPK was observed in the GDNF-treated cells. Functional studies showed that the activation of these MAPKs was critical for Hs683 cell migration induced by GDNF. Our findings revealing molecular mechanisms for the promoting effect of GDNF on glioma cell migration may provide an insight into a better understanding to the malignant progression of human gliomas.