Overexpression of urokinase plasminogen activator (uPA) has been proposed to be one of the mechanisms by which malignant glioma cells cause pericellular proteolysis of stromal structures during brain-tissue invasion. However, knowledge about the mechanisms that regulate uPA in glioma cells is still rather scant. Growth factors, particularly epidermal growth factor and basic fibroblast growth factor (bFGF), regulate uPA synthesis in various nonglial cells. Because these factors have been associated with glioma invasion, we thought that perhaps similar events may occur in glioma cells. In this study, we demonstrate that growth factors regulate uPA gene transcription in glioma cells by induction of Ets-1 transcription factor. Serum and bFGF treatment concomitantly stimulated baseline levels of Ets-1 and uPA mRNA and protein, and this was associated with a marked increase in the migration and invasion potentials of glioma cells in vitro. Treatment of cells with antisense Ets-1 but not the control sense oligonucleotides concurrently inhibited the expression of Ets-1 and uPA, and this blocked glioma cell migration and invasion by more than 50%. In addition, medium conditioned by antisense Ets-1 oligonucleotide-treated cells showed markedly reduced uPA activity, as determined by casein zymography. These results strongly suggest that serum and bFGF control glioma invasion by inducing synthesis of Ets-1 transcription factor, which directly up-regulates expression of uPA in glioma cells. Antisense inhibition of Ets-1 expression may therefore provide a potential and exciting therapeutic target for preventing invasion by glioma cells.