Untargeted LC-MS Metabolomics Reveals Source-Dependent Metabolic Remodeling of Caco-2 Cells Exposed to Bovine Hydatid Cyst Fluid.

Hydatid cyst fluid (HCF) is a biochemically complex parasite-derived mixture whose effects on host and cancer cells are highly context dependent. The aim of this study was to determine whether short-term exposure to bovine liver- or lung-derived HCF alters Caco-2 XTT-based viability/metabolic activity and to characterize the associated global metabolic changes using untargeted liquid chromatography-mass spectrometry (LC-MS). Caco-2 cells were exposed to HCF at 20-60 μg/mL for 24 h for XTT analysis, and 40 μg/mL was selected for untargeted LC-MS metabolomics. XTT-based viability/metabolic activity did not differ significantly from the control across the tested concentration range. In contrast, metabolomic profiling revealed coordinated remodeling of multiple metabolic pathways. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and variable importance in projection (VIP) analysis showed group-specific metabolic signatures and identified metabolites contributing to HCF-associated separation. Changes shared by both HCF sources included increased cytidine, L-valine, N1-acetylspermidine, N6,N6,N6-trimethyl-L-lysine, phenylethylamine, 4'-phosphopantothenoylcysteine, and the signal annotated as reduced riboflavin, together with decreased sphingosine, glucose-6-phosphate, citrate, phenylalanine, methionine, and choline. Source-dependent differences were also observed: lung-derived HCF produced stronger cytidine and L-valine responses, whereas liver-derived HCF was associated with increased glutathione and dolichyl-β-D-glucosyl phosphate. Overall, the 24-h response is consistent with early metabolic adaptation rather than acute cytotoxicity and suggests altered pyrimidine handling, branched-chain amino acid and polyamine metabolism, sphingolipid signaling, flavin cofactor handling, and endoplasmic-reticulum-associated glycosylation. These findings identify bovine HCF as a source-dependent metabolic stressor in colorectal epithelial cancer cells and provide testable targets for future mechanistic validation.