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Objective: To evaluate the cytotoxic,apoptotic,mutagenic and immunomodulatory activities of Kaempferia galanga Linn.(KG) extract and ethyl-p-methoxycinnamate (EPMC) in vitro.Methods: The present study investigated the cytotoxic[using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphe nyl-2-H-tetrazolium bromide test],apoptotic (using a mitochondrial membrane potential assay),muta-genic (using a micronucleus test) and immunomodulatory (using flow cytometry) activities of the ethanolic extract of KG and its bioactive component,EPMC,against two cholangiocarcinoma (CCA) cell lines,CL-6 and HuCCT1,and one normal human cell line,OUMS-36T-1F.Results: Both KG extract and EPMC exhibited moderate cytotoxic activity against both CCA cells.The cytotoxic activity was supported by their concentration-dependent induction of apoptosis.CL-6 was most sensitive (3-4 fold) and selective to 5-fluorouracil (5-FU),compared with KG extract and EPMC[median half inhibiting concentration (ICso) and selectivity index (SI) were 23.01 μg/mL and 17.32;78.41 μg/mL and 4.44;100.76 μg/mL and 2.20,respectively for 5-FU vs.KG extract vs.EPMC].HuCCT1 was relatively more sensitive and selective to 5-FU and EPMC than KG extract[median ICso and SI were 66.03 μg/mL and 6.04;60.90 μg/mL and 3.65;156.60 μg/mL and 2.23,respectively for 5-FU vs.EPMC vs.KG extract].EPMC produced relatively potent cytotoxic activity against polymorphonuclear cells (IC5o =92.20 μg/mL).KG extract and EPMC exhibited concentration-dependent mutagenic activity,as well as inhibition of tumor necrosis factor-α and interleukin-6.Conclusion: Considering cytotoxic,apoptotic,immunomodulatory and mutagenic activities,further development of KG as a drug candidate is likely to focus on the oral pharmaceutical formulation of a standardized KG extract rather than isolated compounds.