Previous studies have confirmed the role of phosphorylated form of 4E-binding protein 1 (p-4E-BP1) as a good candidate tumor biomarker. The aim of this study was to investigate p-4E-BP1 expression status in hilar cholangiocarcinoma (HCCA) specimens and to clarify its clinical significance. Tissue microarray containing tumor specimens obtained from 61 patients with HCCA were constructed. p-4E-BP1 was investigated by immunohistochemical studies. High/moderate expression p-4E-BP1 was observed in 57.4% (35/61) primary cancer specimens. Overexpression of p-4E-BP1 protein was associated with poor differentiation and regional lymph node metastasis. Survival analysis and Cox proportional hazards model revealed that p-4E-BP1 overexpression was an independent factor in predicting recurrence-free survival and overall survival for HCCA patients, apart from tumor invasion and complete resection. P-4E-BP1 was highly expressed in HCCA. Overexpressed p-4E-BP1 might be a novel biomarker to predict the clinical outcome of patients with resected HCCA.