To further get the evidences that hirudin regulates autophagy procession, the three glioma cell lines were treated with hirudin or in combination with the inhibitors for early‐ or late‐stage autophagy, including 3‐methyladenine (3‐MA), Bafilomycin A1 (BafA1) or chloroquine (CQ) and following western blot analysis was performed to examine the changes of LC3‐II and p62/SQSTM1, an indicator of the late stage autophagic flux which is normally degraded by the lysosomal proteases through the interaction with LC3‐II in the late stage of autophagy. This evidence concerns the gene SQSTM1 and central nervous system cancer.