Objective To investigate the effect of heparanase（HPSE）on invasion and migration of gall⁃ bladder cancer cells and ERK/MMP⁃9 signaling pathway. Methods Lentivirus transfection was used to bidirectional regulate the expression of HPSE gene in gallbladder cancer cells（GBC ⁃ SD）to obtain a transgenic gallbladder cancer cell line with overexpression of HPSE and a gallbladder cancer cell line with very low expression of HPSE. The overexpression and interference effects of HPSE were detected by qRT⁃PCR and Western blot. Transwell and cell scratch test were used to detect the invasion and migration ability of gallbladder cancer cells in negative control group，HPSE knockdown group and HPSE overexpression group before and after transfection. At the same time，the mRNA transcription levels of HPSE and MMP ⁃9 were detected by qRT ⁃PCR，and the protein expression levels of MMP⁃9，HPSE，p⁃ERK，ERK，N⁃cadherin and E⁃cadherin were detected by Western blot. Results The gallbladder cancer cell lines of HPSE gene overexpression group and interference group were successfully constructed（P < 0.05）. Overexpression of HPSE significantly promoted the migration and invasion of gallbladder cancer cells，and the expression of MMP⁃9 mRNA and protein，p⁃ERK and N⁃cadherin protein was up⁃regulated，while the expression of E⁃cadherin was down⁃regulated. After knockdown of HPSE expression，the situation was opposite（P < 0.05）. In addition，the co ⁃treatment of HPSE overexpression and ERK inhibitor reversed the promotion effect of HPSE gene overexpression on the biological function of gallbladder cancer cells（P < 0.05）. Conclusions Heparanase enhances the invasion and migration of gallbladder cancer cells by activating ERK signaling pathway and upregulating MMP⁃9.