OBJECTIVE To identify potential cell signaling pathways and protein targets of the active compound isolated from Atracylodes lancea "atractylodin" in cholangiocarcinoma, using proteomics approach. METHODS The holangiocar- cinoma cell line was exposed with atractylodin for 3 and 6 h and the proteins from both intra- and extra- cellular components were extracted. The extract proteins were separated by SDS-PAGE and digested with trypsin.The LC-MS/MS was applied to identify proteins. Signaling pathways and protein expression were analyzed by MASCOT and STITCH software.RESULTS A total of 4,323 and 4,318 proteins were identified from intra-and extracellular components,respectively. Six intracellular proteins were linked with the signaling pathways (apoptosis, cell cycle control, and PI3K-AKT).Four extracellular proteins were linked with the signaling pathways(NF-κB and PI3K-AKT). CONCLUSION All these proteins will further study to confirm the link to the anticholangiocarcinoma ac-tivity of actractylodin.