[Ursolic acid in Formula inhibits colorectal cancer cell growth by inducing cuproptosis].

[OBJECTIVES] To investigate the molecular mechanism by which Formula (HJF) and its active component ursolic acid inhibit colorectal cancer (CRC) cell growth.

[METHODS] Proteomics was used to analyze the effect of HJF on protein expression profile in CRC xenografts from tumor-bearing nude mice. Serum pharmacochemistry was used to identify the potential active components of HJF. Network pharmacology and molecular docking were employed to predict the interaction between ursolic acid and cuproptosis-related targets. Cellular assays including MTT, wound healing, colony formation, and Western blotting were used to validate the effects of ursolic acid on proliferation, migration, and cuproptosis-related indicators (FDX1, SLC31A1, DLAT, GSH, MDA, pyruvic acid, and Cu²⁺) in HCT-116 and LoVo cells.

[RESULTS] HJF regulated 628 differentially expressed proteins in CRC, involving pathways related to inflammation, immunity, and metabolism. Ursolic acid was identified as a major blood component of HJF and exhibited a strong binding affinity with the key cuproptosis protein FDX1 (LiDock Score106.813). In HCT-116 and LoVo cells, ursolic acid significantly inhibited cell proliferation and migration, induced intracellular accumulation of Cu²⁺, MDA and pyruvic acid, reduced GSH levels, inhibited cellular DLAT expression, and up-regulated the expressions of FDX1 and SLC31A1.

[CONCLUSIONS] As one of the key active components in the HJF, ursolic acid inhibits CRC cell growth by inducing cuproptosis targeting FDX1.