Objective: To study the significance of p28GANK expression in hilar cholangiocarcinoma and its relation with the postoperative prognosis of the patients. Methods: The p28GANK expressions in hilar cholangiocarcinoma tissues (62 cases) and normal bile duct tissues (16 cases) were determined by immunohistochemical staining, and the relations of p28GANK expression level with the clinicopathologic parameters and postoperative survival of the patients were analyzed. Results: The p28GANK positive expression rate in cholangiocarcinoma tissues was 74.2% (46/62), while in normal bile duct tissues there was no expression, and the difference between them had statistical significance (P<0.01). The p28GANK expression levels in cholangiocarcinoma tissue showed no statistical difference between the groups divided by age or sex (both P>0.05), but were statistically different between or among the groups stratified by degree of tumor differentiation, TNM stage, portal vein invasion, and lymph node metastasis (all P<0.05). The 5-year overall survival rate of p28GANK positive expression patients was significantly lower than that of p28GANK negative expression patients (P=0.001). Conclusion: p28GANK expression is increased in hilar cholangiocarcinoma, and its high expression is closely associated with the degree of malignant progression of the tumor and also indicates a poor prognosis.