PURPOSE To study the expression of breast cancer metastasis suppressor 1 (BRMS1) and heparanase (HPA) and evaluate their clinicopathologic significance and relationship in benign and malignant lesions of the gallbladder. MATERIALS AND METHODS EnVision immunohistochemical method for determining the expression of BRMS1 and HPA was used in routinely paraffin-embedded sections of surgical resected specimens from gallbladder adenocarcinoma, peritumoral tissues, polyp, and chronic cholecystitis. RESULTS The positive rate of BRMS1 expression was significantly lower in gallbladder adenocarcinoma than that in peritumoral tissues (P<0.01), polyp (P<0.01), and chronic cholecystitis (P<0.01). The positive rate of HPA expression was significantly higher in gallbladder adenocarcinoma than that in peritumoral tissues (P<0.01), polyp (P<0.05), and chronic cholecystitis (P<0.01). The positive expression of BRMS1 and negative expression of HPA were significantly associated with differentiation, lymph node metastasis, and invasion of adenocarcinoma (P<0.05 or P<0.01). Unitivariate Kaplan-Meier analysis showed that decreased expression of BRMS1 (P=0.008) or increased expression of HPA (P=0.016) was associated with decreased overall survival. Multivariate Cox regression analysis showed that decreased expression of BRMS1 (P=0.011) and/or increased expression of HPA (P=0.019) was an independent bad prognostic predictor in gallbladder adenocarcinoma. CONCLUSION The expression of BRMS1 and/or HPA might be closely related to the carcinogenesis, clinical biological behaviors, and prognosis of gallbladder adenocarcinoma.