Objective:To study the roles of hepatoma-derived growth factor (HDGF), vascular endothelial growth factor (VEGF) and mucin 5B (MUC5B) in hepatolithiasis associated with intrahepatic cholangiocarcinoma (HICC).Methods:The clinical data and tissue specimens of 116 patients who underwent hepatectomy at the Third Affiliated Hospital of Wenzhou Medical University from October 1999 to October 2019 were retrospectively analyzed. There were 41 patients with HICC (the HICC group), 38 patients with intrahepatic cholangiolithiasis (the intrahepatic cholangiolithiasis group), and 37 patients with benign liver tumor who underwent hepatectomy (the control group). There were 47 males and 69 females, with age of (66.1±3.2) years old. The positive expressions of HDGF, VEGF and MUC5B in the three groups were compared. In 41 patients with HICC, the correlation between positive expressions of HDGF, VEGF and MUC5B with patients’ clinical characteristics were studied.Results:Compared with the control group, the positive expression rates of HDGF, VEGF and MUC5B in the HICC group and the intrahepatic cholangiolithiasis group were significantly increased, ( P<0.05). Compared with the intrahepatic cholangiolithiasis group, the positive expression rates of HDGF, VEGF and MUC5B in the HICC group were significantly increased ( P<0.05). Positive expression of VEGF in HICC patients was correlated with tumor differentiation, tumor local invasion, tumor length, tumor stage, tumor carbohydrate antigen CA19-9 level and lymph node metastasis ( P<0.05). Spearman correlation analysis showed that HDGF was positively correlated with VEGF expression in HICC tissues specimens ( r=0.935, P<0.01). Survival analysis showed that the cumulative survival rates (36.7%, 17.1%, 7.3%) of patients with positive expression of VEGF were significantly lower than that of patients with negative expression (51.2%, 26.8%, 19.5%) at 1, 3, 5 years after surgery ( P<0.01). The cumulative survival rate (34.1%, 17.1%, 4.9%) of patients with MUC5B positive expression were significantly lower than that of patients with negative expression (53.7%, 31.7%, 17.1%) at 1, 3, 5 years after surgery ( P<0.01). Conclusion:HDGF can be used as a reference indicator for early assessment of HICC. Overexpressions of VEGF and MUC5B can be used as important indicators for HICC in evaluating disease progression and prognosis.