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Apoptosis is an essential type of programmed cell death. Previous studies have demonstrated that a wide range of natural-derived anticancer agents induce apoptosis by trigging oxidative stress. Artemisia argyi is a traditional Chinese herb for treating diverse diseases including dyspepsia, arthroncus, and anaphylactic disease. In this study, sesquiterpene lactone 3 (SL3), a bioactive ingredient isolated from Artemisia argyi was found to show obvious inhibitory effect on two gastric carcinoma cells. Mechanism study revealed that SL3 promoted the membrane translocation of p47, activated nicotinamide adenine dinucleotide (NADPH) oxidase, and evaluated intracellular reactive oxygen species production, leading to the activation of mitochondria-dependent caspase apoptosis pathway. Collectively, these findings show that SL3 is a promising anticancer candidate against gastric carcinoma by activating NADPH oxidase/reactive oxygen species/mitochondrial pathway.