Abstract: Objective To investigate the effect of cathepsin B on invasion and metastasis of laryngeal cancer cells. Methods Hep-2 cells were treated with Cathepsin B specific inhibitor CA-074Me.Then,the expression of Cathepsin B protein was detected by Western blot,the invasion and migration ability of Hep-2 cells was detected by Transwell assay,the proliferation ability of Hep-2 cells was detected by MTT assay,and the growth curve was drawn. Results At 24 hours after CA-074Me treatment to Hep-2 cells,the expression of Cathepsin B protein significantly decreased (P<0.05).Transwell test showed that the invasion and migration ability of Hep-2 cells in vitro was significantly lower than that of control group (P<0.05).MTT assay showed that the proliferation of laryngeal cancer cells in the experimental group significantly reduced. Conclusion Inhibiting the expression of Cathepsin B can significantly reduce the proliferation,invasion and migration ability of tumor cells in vitro,which suggests that Cathepsin B may be regarded as a therapeutic target for laryngeal cancer.