Crocin Promotes Ferroptosis in Gastric Cancer via the Nrf2/GGTLC2 Pathway

Nan Yan ^1,2,3,4 Wei Zhou ⁵ Gaofu Li ⁵ Linglin Zhao ^2,3,4,6 Qijing Guo ⁷ Jie Yang ⁷ Jianhong Liu ^8* Yue Gao ^5* Yushuang Luo ^1,2,4,7,9*

• ¹ Research Center for High Altitude Medicine, Qinghai University, Xining, Qinghai Province, China
• ² Laboratory of High Altitude Medicine in Qinghai Provincial, Qinghai University, Xining, Qinghai Province, China
• ³ Key Laboratory of High Altitude Medicine (Ministry of Education), Qinghai University, Xining, Qinghai Province, China
• ⁴ Key Laboratory of Application and Foundation for High Altitude Medicine Research in Qinhai Province (Qinghai-Utah Joint Research Key Lab for High Altitude Medicine), Qinghai University, Xining, Qinghai Province, China
• ⁵ Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, Beijing, China
• ⁶ Research Center for High Altitude Medicine, Qinghai University Medical College, Xining, Qinghai Province, China
• ⁷ Affiliated Hospital of Qinghai University, Xining, Qinghai Province, China
• ⁸ College of Humanities and Technology, Qinghai Open University, xining, China
• ⁹ Key Laboratory of High Altitude Medicine (Ministry of Education), Xining, Qinghai Province, China

Gastric cancer (GC) is characterized by a high incidence and a low survival rate. The bioactive compound crocin, found in saffron, has demonstrated anti-cancer properties; however, its mechanisms in GC remain unclear. This study investigated the effects of crocin on the proliferation, apoptosis, migration, invasion, and ferroptosis of GC cells using both in vivo and in vitro. We analyzed differentially expressed genes and conducted pathway enrichment analysis following crocin treatment, as well as explored its mechanisms through molecular docking and ChIP-qPCR. The results indicated that crocin inhibited the proliferation, migration, and invasion of GC cells while promoting apoptosis. After crocin treatment, differentially expressed genes were primarily enriched in pathways related to oxidative stress and ferroptosis. Crocin inhibited GC proliferation, invasion, and migration by downregulating the oncogene GGTLC2, while simultaneously promoting apoptosis and ferroptosis. Molecular docking results indicate that crocin has a stable binding affinity for GGTLC2, suggesting that it may directly target GGTLC2 to regulate its expression. Additionally, crocin facilitated the translocation of the transcription factor Nrf2 from the nucleus to the cytoplasm. ChIP-qPCR results indicated that Nrf2 can directly bind to the promoter region of GGTLC2 to regulate its expression, and crocin inhibited this binding effect. Therefore, we propose that crocin, as a promising herbal monomer for GC treatment, may inhibit ferroptosis in GC cells through the Nrf2/GGTLC2 pathway, thereby suppressing the occurrence and progression of GC.