Glucose-6-phosphate dehydrogenase (G6PD) shields pancreatic cancer from autophagy-dependent ferroptosis by suppressing redox imbalance induced AMPK/mTOR signaling.

Pancreatic cancer is a highly aggressive malignancy with a significant unmet medical need, as current treatments often yield poor responses. Ferroptosis, a recently recognized form of regulated cell death, has garnered increasing attention for its potential in cancer therapy. However, the molecular links connecting autophagy to ferroptosis remain largely unclear. In this study, we identified that the redox related protein glucose-6-phosphate dehydrogenase (G6PD) is overexpressed in pancreatic cancer and correlates with poor prognosis, promoting cancer cell proliferation and migration. Using PANC-1 and MiaPaCa-2 pancreatic cancer cell lines, we demonstrated that the treatment with ferroptosis-inducing compound RSL3 induced glycolytic dysfunction and significantly downregulated G6PD expression. Moreover, G6PD knockdown in these cell lines impaired the cellular antioxidant defence capability by decreasing the NADPH and GSH contents, leading to increased lipid peroxidation and malondialdehyde (MDA) accumulation. Particularly, G6PD depletion exacerbated RSL3 induced oxidative stress and synergistically promoted autophagy-dependent ferroptosis. Mechanistically, we found that G6PD knockdown disrupted redox homeostasis, triggering the activation of AMPK-mTOR pathway to induce autophagy. Furthermore, pharmacological inhibition of AMPK (with Compound C) rescued ferroptosis induced by G6PD knockdown and RSL3, whereas mTOR inhibition (with Rapamycin) further augmented cell death. Altogether, these findings suggest that G6PD contributes to ferroptosis resistance in pancreatic cancer cells by modulating oxidative balance and autophagy via the AMPK-mTOR pathway, highlighting its potential as a therapeutic target.