ackground: Chemotherapeutic treatment ofcholangiocarcinoma is largely ineffective, because it isdue to non-responsive for the obscured reason of thecancer to anticancer agents. Study was to investigate thecytotoxicity effects of chemotherapeutic agents andcurcumin in vitro.Objective: To test sensitivity of cholangiocarcinoma celllines to chemotherapeutic agents and curcuminMaterial and Methods: Three cholangiocarcinoma (CCA)cell lines, including KKU-100, KKU-M214 and KKU-OCA17were used in the study. All cell lines were treated with threechemotherapeutic agents (5-fluorouracil, doxorubicin andcarboplatin) or curcumin. The cell viability was determinedunder a fluorescence microscope by counting the numberof living and dead cells after treatment.Results: KKU-100 and KKU-M214 were the very sensitivecell lines to doxorubicin (IC50 values ranging from 0.4 to0.7 nmol/l), whereas KKU-OCA17 was relatively moreresistant cell line. Interestingly, all cell lines were sensitiveto curcumin (IC50 values ranging from 3 to 17 μmol/l).Conclusion: This study showed different degrees ofsemsitivity of CCA cell lines to various chemotherapeuticagents. The data could serve as a basic information forchemotherapeutic selection for the treatment in CCApatients. Moreover curcumin exhibits the cytotoxicity onCCA cell lines, thereby it is suggested to be very beneficialin the development of strategy for chemotherapy of CCA cancer.