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13–15 Calycosin is a fat-soluble component in the dried root of Astragalus membranaceus, which are organic compounds widely found in nature. Calycosin exerts various anti-tumor activities in vitro and in vivo, 16–18 but its mechanism of action in human GC cells is not clear. In this study, we evaluated the effects of calycosin on apoptosis, the cell cycle, and migration of GC cells under ROS regulation. In addition, we also determined the roles of MAPK/STAT3/NF-κB pathways in these processes. Materials and Methods Cell Lines and Cell Culture Twelve types of GC cells (AGS, KATO-3, MKN-28, MKN-45, NCI-N87, SNU-5, SNU-216, SNU-484, SNU-668, YCC-1, YCC-6, and YCC-16) and normal human lung cells IMR-90 were purchased from American Type Culture Collection (Manassas, VA, USA), and three normal cell lines (GES-1, L-02, and 293T) were purchased from Saiqi Biotech Co., Ltd (Shanghai, China). AGS, KATO-3, MKN-28, MKN-45, SNU-5, SNU-216, SNU-484, SNU-668, IMR-90, ....