Abstract
Abstract As a common malignant tumor, achieving better treatment outcomes for liver cancer remains our constant pursuit. To enhance tumor cell killing efficiency, this study designed a nanozyme—gambogic acid-iron-doxorubicin (GAFe@Dox)—that simultaneously improved the tumor microenvironment (TME) and sensitized cells to chemotherapy. Its antitumor efficacy was systematically evaluated using 3D hydrogel cell scaffolds model simulating the human TME. First, transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS) confirmed the successful synthesis of GAFe@Dox and verified its peroxidase activity. Cellular experiments demonstrated that GAFe@Dox generated substantial reactive oxygen species (ROS) via the Fenton reaction, synergistically killing tumor cells through multiple pathways with other components. Its killing mechanism was preliminarily validated at the cellular level. Transcriptomic analysis revealed GAFe@Dox effectively regulated signaling pathways including p53, MAPK and so on, further substantiating its mechanism of action. Additionally, we successfully constructed and cultured 3D hydrogel cell scaffolds model, simulating the in vivo 3D environment for cell growth and proliferation, achieving preliminary success. Subsequent validation confirmed that GAFe@Dox effectively penetrated and inhibited cell proliferation while inducing tumor cell apoptosis. This study demonstrated GAFe@Dox’s potent cytotoxic effects through synergistic multi-mechanistic tumor cell killing and established a physiologically relevant drug efficacy evaluation model, offering novel insights for nanozymes therapy research in liver cancer.