Exploring the mechanism of cinobufagin on gastric cancer based on proteomics, molecular dynamics, in vitro and in vivo experiment

Cinobufagin as one of the primary bioactive components of Chansu, which is a widely used traditional remedy in China, has been employed in the treatment of various malignant tumors, including gastric cancer. However, its antitumor effects on gastric cancer and the underlying mechanisms have not yet been fully elucidated. This study aims to comprehensively investigate the effects of cinobufagin on AGS gastric cancer cells both in vitro and in vivo, and to further explore its mechanism of action. AGS gastric cancer cells were treated with cinobufagin. A series of assays and analyses were conducted, and an in vivo gastric cancer model, to examine changes in AGS cell characteristics and to elucidate the molecular mechanisms underlying the effects of cinobufagin. Our data demonstrate that cinobufagin exhibits multiple anti-gastric cancer activities, including suppression of proliferation, induction of apoptosis, cell cycle arresting at the G0/G1 phase, and inhibition of migration and invasion in AGS gastric cancer cells. Furthermore, proteomic analysis, bioinformatics analysis, molecular docking, and molecular dynamics simulation suggest that decorin (DCN) is the most probable target of cinobufagin. Western blot and immunohistochemistry (IHC) assays confirmed that cinobufagin significantly upregulated DCN expression in both AGS cells and xenograft tumors. Moreover, cinobufagin effectively reduced the levels of EGFR and active EGFR, and inhibited Erk phosphorylation, which may contribute to its anti-gastric cancer effects. Cinobufagin is capable of suppressing gastric tumor growth in mice likely through the DCN/EGFR pathway and may serve as a potential treatment option for gastric cancer.