Identification of ferroptosis as a novel mechanism for antitumor activity of natural product derivative a2 in gastric cancer

Ferroptosis is a type of cell death accompanied by iron-dependent lipid peroxidation, thus stimulating Ferroptosis may be a potential strategy for treating gastric Cancer, therapeutic agents against which are urgently required. Jiyuan oridonin A (JDA) is a natural compound isolated from Jiyuan Rabdosia rubescens with anti-tumor activity, unclear anti-tumor mechanisms and limited water solubility hamper its clinical application. Here, we showed a2, a new JDA derivative, inhibited the growth of gastric Cancer cells. Subsequently, we discovered for the first time that a2 induced Ferroptosis. Importantly, compound a2 decreased GPX4 expression and overexpressing GPX4 antagonized the anti-proliferative activity of a2. Furthermore, we demonstrated that a2 caused ferrous iron accumulation through the Autophagy pathway, prevention of which rescued a2 induced ferrous iron elevation and cell growth inhibition. Moreover, a2 exhibited more potent anti-cancer activity than 5-fluorouracil in gastric Cancer cell line-derived xenograft mice models. Patient-derived tumor xenograft models from different patients displayed varied sensitivity to a2, and GPX4 downregulation indicated the sensitivity of tumors to a2. Finally, a2 exhibited well pharmacokinetic characteristics. Overall, our data suggest that inducing Ferroptosis is the major mechanism mediating anti-tumor activity of a2, and a2 will hopefully serve as a promising compound for gastric Cancer treatment.

5-FU, 5-fluorouracil; Autophagy; CDX, cell line-derived xenograft; DCFH-DA, dichlorodihydro-fluorescein diacetate; DCM, dichloromethane; Ferroptosis; Ferrous iron; GPX4; Gastric cancer; IKE, imidazole ketone erastin; JDA derivative; JDA, Jiyuan oridonin A; Jiyuan Rabdosia rubescens; KEGG, Kyoto Encyclopedia of Genes and Genomes; NAC, N-acetylcysteine; PARP, poly ADP-ribose polymerase; PDX, patient-derived tumor xenograft; PK, pharmacokinetic; Papp, apparent permeability coefficient; ROS; ROS, reactive oxygen species; RTV, relative tumor volume; Verp, verapamil; qRT-PCR, quantitative real time PCR.