Abstract: Objective To investigate the molecular mechanism of the induction of gastric cancer cell line AGS death by p97 inhibitor eeyarestatinⅠ (EerⅠ). Methods AGS cells were treated with EerⅠ. Then liquid chromatography-mass spectrometry was used to perform proteome analysis for screening differentially expressed proteins and to find underlying signaling pathways. At the same time, the proteins of related pathway were investigated by protein immunoblotting. Cell proliferation was detected using the CCK-8 test kit; Cell apoptosis was detected using TUNEL staining test kit; Liperfluo probe was used to detect ferroptosis-related lipid peroxides. Results In EerⅠ treatment group, there were significant changes in proteins(fold change＞1.5 and P＜0.05), in which 125 proteins were increased and 132 proteins were decreased. The enrichment analysis of these DEPs showed that EerⅠ might significantly affect cell apoptosis and ferroptosis-related signaling pathways. Furthermore, EerⅠ could increase genomic DNA fragmentation related to cell apoptosis, increase of lipid peroxides in the ferroptosis pathway, causes changes in cell death related proteins, and inhibit the proliferation of gastric cancer cells. Conclusions p97 inhibitor EerⅠ can induce cell apoptosis and ferroptosis in AGS cells, thereby inhibiting tumor cell proliferation.

Key words: gastric cancer, p97 protein, cell apoptosis, ferroptosis