Objective To study the relation between retinoblastoma (Rb) gene expression and biological characteristics of gallbladder carcinoma. Methods The expression of Rb protein in tissues of 41 cases of gallbladder carcinoma, 7 cases of gallbladder papilloma, and 14 cases of cholecystitis were detected by immunohistochemical staining of SP with polyclonal antibody. Results The rate of positive staining was 58.7% in gallbladder carcinoma which was significantly lower than that in cholecystitis and gallbladder papilloma (100%). There was a significant difference of Rb protein expression among the low, moderate and high differentiations and so was between S5, S4, S1, S2 and S3 stage (P<0.05). Conclusion It sugests that expression or non-expression of Rb gene is closely related to the malignant potential invasiveness and prognosis of gallbladder carcinoma.