Objective: To investigate Maspin protein expression in cholangiocarinoma tissue as well as its clinical significance and relation with p53 protein. Methods: The Maspin and p53 protein expressions in 42 specimens of cholangiocarinoma tissues along with their adjacent tissues and 12 specimens of normal biliary duct tissues were detected by immunohistochemcal staining. The relations of the two proteins with clinicopathologic factors and prognosis of the patients and the relationship between the two proteins were analyzed. Results: The positive expression rates of both Maspin and p53 proteins in cholangiocarinoma tissue were significantly higher than those in cancer adjacent tissue and normal bile duct tissue (all P<0.05); the Maspin protein expression was significantly related to lymph node metastasis and distant metastasis of cholangiocarinoma, while p53 protein expression was significantly associated with the pathological grade, TNM stage, lymph node metastasis and distant metastasis of cholangiocarinoma, but there was no significant correlation between Maspin and p53 protein expression in cholangiocarinoma tissue (r=–0.329, P=0.144). The postoperative survival time in patients with positive Maspin expression was significantly longer than those with negative Maspin expression (χ2=4.440, P=0.035), while in patients with positive p53 expression was significantly shorter than those with negative p53 expression (χ2=8.231, P=0.004). Results of univariate and multivariate Cox regression analysis showed that Maspin protein expression was an independent prognostic index for holangiocarcinoma patients (HR=0.094, 95% CI=0.015–0.593, P=0.012). Conclusion: The Maspin protein expression is increased in cholangiocarcinoma tissue, but its deficiency or absence may closely be related to unfavorable clinicopathologic features and outcomes of the patients, and the mechanism of its action may not be associated with p53 protein.