The modulatory effects of genistein and vitamin K 1 on ghrelin and viability in Caco-2 cells: An in vitro and in silico study

Objective This study evaluated the effects of genistein, alone and combined with vitamin K 1 , on cell viability/metabolic activity and supernatant ghrelin concentrations in Caco-2 cells and explored potentially relevant molecular pathways. Materials and Methods Cell viability and metabolic activity were assessed using trypan blue exclusion and MTT assays, respectively, and ghrelin concentrations in culture supernatants were measured by ELISA. Molecular docking was used to explore potential interactions with GHSR1α, whereas gene-expression and network pharmacology analyses were performed independently to examine colon cancer-related molecular pathways. Results Genistein reduced the viability of Caco-2 cells at high concentrations. However, vitamin K 1 alone did not show a significant effect. In the combination treatment, the MTT signal decreased at the 24th and 48th hours at the selected doses, while no significant difference was observed at the 72nd hour. ELISA results showed that ghrelin concentrations in the supernatant were lower at the 72-h mark. Docking analysis revealed potential interactions of these dietary compounds with GHSR1α, while network analyses identified the PI3K-Akt and MAPK/ERK pathways associated with RTK. Conclusion Combined genistein and vitamin K 1 treatment produced time-dependent biological effects in Caco-2 cells. The accompanying in silico analyses suggest potentially relevant ghrelin- and RTK-associated molecular pathways that warrant further mechanistic investigation.