Abstract: Objective To explore the expressions of Annexin A4（ANXA4）and Annexin A6（ANXA6）protein in hilarcholangiocarcinoma（HC）and to determine their relationships with clinical parameters and prognosis. Methods The expressions of ANXA4 and ANXA6 were detected by tissue microarray and immunohistochemistry in 49 HC tissues and 10 tumor adjacent tissues from 2005 to 2007.The relationships between the expressions of ANXA4 and ANXA6 and clinical parameters were analyzed，as well as prognosis. Results The positive rates of ANXA4 and ANXA6 in HC tissues（63.3％，69.4％）were significantly higher than those（20.0％，30.0％）in tumor adjacent tissues（P＜0.05）. The expression of ANXA4 was closely associated with lymph node metastasis（P＜0.05）. The expression of ANXA6 was closely associated with tumor size，lymph node metastasis and TNM stage（P＜0.05）. The median overall survival（OS）and progression-free survival（PFS）of patients with ANXA4 positive expression was 14 months and 12 months，shorter than 46 months and 39 months of patients with ANXA4 negative expression（P＜0. 05）. The median OS and PFS of patients with ANXA6 positive expression was 14 months and 12 months，shorter than 42 months and 39 months of patients with ANXA6 negative expression（P＜0.05）. Multivariate analysis revealed that ANXA6 and invasion depth were independent prognostic factors influencing OS，and invasion depth was also an independent factor influencing PFS. Conclusion ANXA4 and ANXA6 are associated with occurrence and development of HC，and ANXA6 may be a new tumor marker for predicting prognosis of HC.