Abstract: Objective To explore the risk factors of hepatolithasis-associated intrahepatic cholangiocarcinoma (HICC) and the clinical value of serum tumor-related markers for the detection of HICC. Methods Clinical data were collected from 58 patients pathologically diagnosed as HICC between 2005 and 2011 in West China Hospital of Sichuan University and 189 patients diagnosed as hepatolithiasis alone in the same period as matched control group. Logistic regression analysis was used to detect the independent risk factors of HICC and ROCs curve were constructed to assess the diagnostic value of CA199, CEA, GGT and ALP. Results The patients in both HICC group and control group presented similar clinical symptoms except weight loss. The results of univariate analysis suggested cholangioenterostomy (P<0.001), early stone removal (OR=0.001), family history of cancer (P=0.001) were associated with the incidence of HICC. The results of Multivariate analysis suggested diabetes mellitus (OR=3.621, 95%CI: 1.333-9.834, P=0.012), family history of cancer (OR=16.830, 95%CI:1.937-146.21, P=0.010), cholangioenterostomy (OR=5.115，95%CI: 1.733-15.098, P=0.003), early removal of stone (OR=0.315, 95%CI: 0.128-0.771, P=0.011) and CA199>100 IU/mL (OR=5.478, 95%CI: 2.539-11.820, P<0.001) were independent risk factors for hICC. Serum CA199 and CEA level presented low diagnostic accuracy, a combined test (CA199>100 IU/mL or CEA>5 ng/mL) showed better diagnostic performance with a 71.05% of sensitivity and 82.05% of specificity. Conclusion Cholangioenterostomy, diabetes, early and complete stone removal were independent risk factors for hepatolithiasis-associated ICC. A combined test of CA199 and CEA could be an effective detecting tool for HICC.