Celastrol Targets FANCD2 to Induce Autophagy-Dependent Ferroptosis in Hepatocellular Carcinoma Cells

Celastrol has been shown to inhibit hepatocellular carcinoma (HCC) progression, but the underlying mechanism is unknown. Fanconi anemia complementation group D2 (FANCD2), a ferroptosis inhibitor, promotes HCC proliferation and invasion. This study aims to investigate whether Celastrol exerts its effects by targeting FANCD2. Using data from The Cancer Genome Atlas (TCGA), we identified differentially expressed genes in HCC utilizing Gene Expression Profiling Interactive Analysis 2 (GEPIA 2). FANCD2 and Celastrol were analyzed for molecular docking using Autodock, which was based on geometric matching and energy matching. The correlation between FANCD2 and survival rate was analyzed using Kaplan-Meier's estimates by log-rank (Mantel-Cox) test. HCC cell lines (SNU-423 and SNU-387) were overexpressed or silenced with FANCD2 and treated with Celastrol. Autophagy and ferroptosis were evaluated by measuring oxidative stress and related markers, and cell function experiments were performed. High expression of FANCD2 was correlated with poor survival in HCC patients. Celastrol targeted FANCD2, reducing its level in SNU-423 and SNU-387 cells. FANCD2 overexpression resulted in increased SNU-423 cell viability, migration, invasion, and tube formation ability, as well as attenuated autophagy and ferroptosis, while FANCD2 knockdown in SNU-387 cells showed opposite effects. Additionally, FANCD2 overexpression reversed the ability of Celastrol to induce autophagy and ferroptosis and to inhibit SNU-423 cell survival in vitro, while FANCD2 knockdown enhanced the effects of Celastrol in SNU-387 cells. Celastrol inhibits malignant behavior in HCC cells by targeting FANCD2 to induce autophagy-dependent ferroptosis.