This study investigated the differential expression of fascin, E-cadherin and vimentin in human carcinogenesis.This study detected their expressions in 142 cholangiocarcinoma and 20 benign bile duct tissue specimens using immunohistochemistry. Clinicopathologic characteristics and survival data were also collected and analyzed for their association with expression of these 3 proteins.The data showed that both fascin and vimentin proteins were significantly overexpressed in cholangiocarcinoma, whereas E-cadherin expression was reduced in cholangiocarcinoma compared with normal tissues. Fascin protein expression was associated with tumor dedifferentiation, venous invasion and lymph node or distant metastasis. Similarly, vimentin expression was associated with tumor dedifferentiation and venous invasion, as well as hepatitis virus infection. In contrast, loss of E-cadherin expression was associated with tumor dedifferentiation (P < 0.05). Because the functions of these 3 proteins were closely related, the data showed that fascin and E-cadherin protein expression was reversely associated. However, fascin and vimentin protein expression was positively associated with cholangiocarcinoma. Cox multiple regression analysis showed that distant tumor metastasis, tumor differentiation and overexpression of fascin and vimentin proteins were all independent risk factors for prognosis (P < 0.05).The data from the current study demonstrated that overexpression of fascin and vimentin proteins could be further evaluated as biomarkers for the prediction of cholangiocarcinoma patient survival.