Calycosin Induces Gastric Cancer Cell Apoptosis via the ROS-Mediated MAPK/STAT3/NF-κB Pathway

• Published in: OncoTargets and therapy Vol. 14; pp. 2505 - 2517
• Main Authors: Zhang, Yu, Zhang, Jian-Qiang, Zhang, Tong, Xue, Hui, Zuo, Wen-Bo, Li, Yan-Nan, Zhao, Yue, Sun, Geng, Fu, Zhong-Ren, Zhang, Qing, Zhao, Xue, Teng, Yue, Wang, An-Qi, Li, Jia-Zhu, Wang, Ying, Jin, Cheng-Hao
• Format: Journal Article
• Language: English
• Published: New Zealand Taylor & Francis Ltd 01.01.2021; Dove
• Subjects: Acetylcysteine; Apoptosis; Bcl-2 protein; Biotechnology; c-Jun protein; Cancer therapies; Caspase-3; Cell cycle; Cell death; Cell migration; Chemotherapy; Cyclin D1; Cyclin E; Cyclin-dependent kinase 2; Cyclin-dependent kinase 4; Cyclin-dependent kinase inhibitor p27; Cytochrome c; Cytotoxicity; E-cadherin; Enzyme inhibitors; Extracellular signal-regulated kinase; Flow cytometry; G1 phase; Gastric cancer; JNK protein; Kinases; Lymphocytes B; Lymphoma; Morphology; Original Research; Proteins; Reactive oxygen species; Reagents; Traditional Chinese medicine; Transcription factors; Western blotting; Beijing China; United States > US; China; Japan; cell migration; apoptosis; calycosin; human gastric cancer; cell cycle; reactive oxygen species
• ISSN: 1178-6930; 1178-6930
• DOI: 10.2147/OTT.S292388

Calycosin, an active compound in plants, can promote the apoptosis of various cancer cells; however, the mechanism by which it regulates reactive oxygen species (ROS) in gastric cancer (GC) cells remains unclear. In this study, we investigated the effects of calycosin on apoptosis, the cell cycle, and migration in GC cells under ROS regulation. The results of the Cell Counting Kit-8 assay suggested that calycosin had significant cytotoxic effects on 12 gastric cancer cells, but no significant cytotoxic effects on normal cells. Hoechst 33342/propidium iodide (PI) double staining and flow cytometry showed that calycosin had clear pro-apoptotic effects on AGS cells. Western blotting revealed that the expression of cytochrome C and pro-apoptotic proteins B-cell lymphoma 2 (Bcl-2)-associated agonist of cell death (Bad), cleaved (cle)-caspase-3, and cle-poly (ADP-ribose) polymerase gradually increased, and the expression of anti-apoptotic protein Bcl-2 gradually decreased. Calycosin also decreased the expression of extracellular signal-regulated kinase, nuclear factor kappa B (NF-κB), and signal transducer and activator of transcription 3 (STAT3), and increased the phosphorylation levels of p38, c-Jun N-terminal kinase, and inhibitor of NF-κB. In addition, calycosin markedly increased ROS accumulation, and pretreatment with active oxygen scavenger n-acetyl-l-cysteine (NAC) clearly inhibited apoptosis. Calycosin downregulated the cell cycle proteins cyclin-dependent kinase 2 (CDK2), CDK4, CDK6, cyclin D1, and cyclin E; upregulated p21 and p27; and arrested cells in the G0/G1 phase. Similarly, calycosin also downregulated Snail family transcriptional repressor 1, E-cadherin, and β-catenin and inhibited cell migration. However, pretreatment with NAC inhibited the calycosin-induced effects of cycle arrest and migration. In summary, calycosin induces apoptosis via ROS-mediated MAPK/STAT3/NF-κB pathways, thereby exerting its anti-carcinogenic functions in GC cells.