LC-MS/MS-based metabolomics analysis and experimental verification reveal the mechanism of Dahuang Mudan Decoction inhibition with the proliferation of colorectal cancer cells.

This study aims to combine in vitro experiments with cell metabonomics used to explore the anti-colorectal cancer effect of Dahuang Mudan Decoction drug-containing serum. CCK-8, colony formation assay, and wound healing were utilized to evaluate the effects of DHMDD drug-containing serum on the proliferation and migration of human colorectal cancer cells. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting (WB) were used to measure the factors related to the PI3K/AKT signaling pathway. Furthermore, the potential metabolic mechanism of DHMDD was examined using cell metabolomics analysis. CCK8 results show that the IC50 of the drug-containing serum for HCT116 cells is 7.8%. Conducting cloning and scratch assays at this concentration, the results show that the drug-containing serum significantly curtailed the proliferation and migration of cells. WB and qRT-PCR showed that the drug-containing serum could significantly reduce the expression levels of AKT1, PI3K, PIK3R1, and PIK3CA and p-PI3K and p-AKT. The results of cellular metabolomics analysis indicate that the drug-containing serum can regulate linoleic acid metabolism, arachidonic acid metabolism, phenylalanine metabolism, and glycerophosphate metabolism of colorectal cancer cells by affecting 18 endogenous metabolites, thus affecting the occurrence and development of cancer cells. In summary, these findings indicate that drug-containing serum of Dahuang Mudan Decoction exhibits effective anti-colorectal cancer activity, promoting cell apoptosis mediated by the PI3K/AKT signaling pathway and participating in the regulation of pathways such as linoleic acid and lipid metabolism. The results underscore the value of Dahuang Mudan Decoction as an important adjunctive therapy for colorectal cancer, proposing new strategies for its treatment.