Development of a ferroptosis-related signature and identification of NOTCH2 as a novel prognostic biomarker in pancreatic cancer

Siyi Zhang¹Xiaoxuan Li¹Xiang-Xue Li¹Zi-Heng Zhang¹Kai-Hui Zhu^2*Jing Guo^1*

• ¹The Affiliated Hospital of Qingdao University, Qingdao, China
• ²Huangdao District People's Hospital, Qigdao, China

Background: Ferroptosis, a regulated form of iron-dependent cell death, has shown promise as an anti-tumor mechanism. However, its role in pancreatic cancer remains largely unexplored. This study aimed to identify a ferroptosis-related prognostic signature and key biomarkers. Methods: Transcriptomic profiles and clinical data of pancreatic cancer patients were obtained from the GEO and TCGA databases. A prognostic signature was constructed using LASSO and Cox regression analysis. The role of a key gene, NOTCH2, was investigated through somatic mutation, functional enrichment, immune infiltration, and drug sensitivity analysis. In vitro, the expression of NOTCH2 was confirmed by Western blot, and its effects on cell proliferation and migration were assessed using MTT, colony formation, and wound-healing assays. Its involvement in ferroptosis was further investigated by measuring intracellular iron, reactive oxygen species (ROS) and C11-BODIPY. Results: We constructed and validated a ferroptosis-related prognostic signature consisting of NOTCH2, KRT18, and H1-2. Patients in the high-risk group, as defined by this signature, exhibited significantly worse overall survival. A nomogram integrating the risk score and clinical variables demonstrated excellent accuracy in predicting patient prognosis. We identified NOTCH2 as a key biomarker, showing upregulated expression in pancreatic cancer tissues and cell lines, which correlated with poor prognosis and increased infiltration of M2 macrophages. Functionally, knockdown of NOTCH2 in vitro inhibited the proliferation and migration of pancreatic cancer cells while increasing both intracellular iron concentration and lipid peroxidation levels. Conclusion: Our study establishes a ferroptosis-related signature for prognostic prediction in pancreatic cancer and identifies NOTCH2 as a critical prognostic biomarker. NOTCH2 may promote pancreatic cancer progression by suppressing ferroptosis, highlighting it as a potential therapeutic target.