Resveratrol (RSV), a polyphenol, is known to play an important role in inhibiting proliferation and inducing apoptosis of glioma cells. The aim of this study was to explore the mechanism of RSV on U251 cells apoptosis. RSV showed a dose-dependent decrease in U251 cell viability. It could reduce IκB phosphorylation, nuclear P65 protein levels and NF-κB transcriptional activity, which suggested that signaling pathway are involved in RSV-induced apoptosis. In addition, RSV could inhibit miR-21 expression and down-regulation of miR-21 expression could suppress NF-κB activity. Interestingly, over-expression of miR-21 can reverse the effect of RSV on NF-κB activity and apoptosis in U251 cells. These results suggest that RSV can effectively induce apoptosis of U251 cells and modulation of miR-21 possibly contributes to this antitumor action.