RHOJ derived peptide promotes chemosensitivity by inhibiting glutamine metabolism in gastric cancer.

BACKGROUND: Chemoresistance is a cause of chemotherapy failure in gastric cancer (GC) treatment. Recent studies have shown that the dysregulation of glutamine metabolism plays a pivotal role in promoting chemoresistance. While small-molecule inhibitors targeting glutamine metabolism have been investigated, peptide-based compounds have attracted increasing attention because of their high specificity and low toxicity. Endogenous or rationally designed peptides have shown potential for inducing apoptosis, disrupting cancer-related signaling pathways, and overcoming drug resistance in various cancers. However, the potential of functional peptides to target glutamine metabolism and reverse drug resistance in the context of GC has not been thoroughly explored. METHODS: We performed proteomic profiling to identify upregulated proteins in cisplatin-sensitive GC cells, from which peptides were derived for functional screening. A RHOJ-derived peptide (peptide 1) was identified and validated as a candidate chemosensitizer. Untargeted metabolomics, flow cytometry, molecular docking, molecular dynamics simulations, fluorescence imaging, and a subcutaneous xenograft model were used to investigate the mechanism through which peptide 1 modulates GLUL-mediated glutamine metabolism and reverses cisplatin resistance. RESULTS: In this study, we found that glutamine metabolism was enhanced in cisplatin-resistant GC cells and that a peptide which derived from RHOJ (peptide 1) increased the sensitivity of resistant cells to chemotherapy. Molecular docking revealed that this peptide could bind to glutamine synthetase (GLUL), a key enzyme in the glutamine metabolism pathway. Mechanistically, peptide 1 inhibited glutamine production, increased ROS levels, induced DNA damage, and promoted apoptosis in resistant cells, ultimately restoring cisplatin sensitivity both in vitro and in vivo. CONCLUSIONS: Our study demonstrated that glutamine metabolism plays a vital role in the chemoresistance of GC cells and that RHOJ-derived peptide 1 enhances the chemosensitivity of drug-resistant GC cells through targeting GLUL, depleting glutamine, inducing ROS accumulation, and promoting DNA damage. These mechanisms ultimately restores chemosensitivity in drug-resistant cells and highlight peptide 1 as a promising therapeutic strategy for overcoming chemoresistance in GC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12967-026-07731-z.