Network Pharmacology and Integrated Experimental Evidence Demonstrate That Ophiopogonin D Suppresses Hepatocellular Carcinoma Progression via the <scp>UCK2</scp> – <scp>SLC7A11</scp> Axis

ABSTRACT Hepatocellular carcinoma (HCC) treatment faces significant challenges, including high toxicity and drug resistance. Ophiopogonin D, the active compound derived from the medicinal food material Ophiopogon japonicus (Mai Dong), has demonstrated potential anti‐cancer properties. However, its specific mechanism in combating HCC remains unclear. To investigate the anti‐HCC effects and underlying mechanisms of Ophiopogonin D, this study employed in vitro experiments, network pharmacology, and in vivo models. The results revealed that Ophiopogonin D exhibited an IC 50 range of 74.71–78.35 μM against Hep3B, HepG2, and Huh7 cells, with minimal toxicity to normal liver cells (LO2). At concentrations between 10 and 160 μM, it concentration‐dependently inhibited HCC cell proliferation, induced apoptosis, and suppressed migration and invasion. Network pharmacology combined with experimental validation identified UCK2 as a key target. Ophiopogonin D downregulated UCK2 expression, promoted reactive oxygen species (ROS) accumulation in HCC cells, induced ferroptosis by downregulating SLC7A11, disrupted the UCK2‐SLC7A11 interaction, and inhibited the PI3K/AKT pathway. In vivo, Ophiopogonin D dose‐dependently inhibited tumor growth in nude mice xenografts. This study demonstrates that Ophiopogonin D exerts anti‐HCC effects through the UCK2‐SLC7A11 axis, providing experimental evidence and molecular mechanisms to support the development of novel anti‐HCC drugs derived from medicinal foods.