Objective To investigate the inhibitory effect of alantolactone on C6 glioma and its possible mechanism. Methods Rat C6 glioma cell line was cultured in vitro, the effect of AL on the viability of C6 cells was detected by MTT assay; to detect the effect of AL on migration and invasion of C6 cells by scratch test and transwell chamber assay; flow cytometry was used to detect the effect of AL on the induction of apoptosis in C6 cells and the effects of AL on the changes of mitochondrial membrane potential in C6 cells by JC-1 fluorescent probe; western blot was used to detect the expression of related proteins in C6 cells after AL treatment. Results AL significantly inhibits the proliferation of C6 cells in a time and dose dependent manner. After treatment with 24h by AL, the migration and invasion of C6 cells were significantly inhibited, N-cadherin protein was significantly decreased and E-cadherin protein was significantly increased. The number of apoptotic cells increased obviously while the mitochondrial membrane potential decreased significantly. The protein expressions of PARP / cleaved caspase-3 / cleaved caspase-9 were significantly up-regulated. Conclusions AL inhibits the mechanism of action of C6 cells by inhibiting the migration and invasion of the cells through regulating the protein of cadherins and inducing the apoptosis of C6 cells by regulating the cytochrome C / Caspase signaling pathway.