Integrated network pharmacology and metabolomics reveal the action mechanisms of vincristine combined with celastrol against colon cancer.

Colon cancer is associated with a high mortality rate. Vincristine (VCR) is a commonly used chemotherapeutic drug. Celastrol (CEL) is an effective component which exerts inhibitory effects on colon cancer. Combination treatment improves resistance to chemotherapeutic drugs and enhances their efficacy. Therefore, we aimed to explore the molecular mechanisms of VCR combined with CEL in colon cancer treatment. We verified the effects of VCR combined with CEL on the proliferation, cell cycle, and apoptosis of HCT-8 cells. Non-targeted metabolomic techniques were used to analyse the changes in cellular metabolites after administration. Finally, network pharmacology technology was used to screen the potential targets and pathways. VCR combined with CEL had synergistic inhibitory effects on HCT-8 colon cancer cells. Cell metabolomics identified 12 metabolites enriched in metabolic pathways, such as the phenylalanine, tyrosine and tryptophan biosynthesis pathways. Network pharmacology revealed that MAPK1, AKT1, PIK3CB, EGFR, and VEGFA were the key targets. Western blotting revealed that VCR combined with CEL activated the P53 pathway by suppressing the PI3K/AKT signalling pathway activation and Bcl-2 expression, promoting the Bax expression. Therefore, VCR combined with CEL potentially treats colon cancer by increasing the apoptosis, improving energy metabolism, and inhibiting PI3K/AKT pathway in colon cancer cells.

[Overexpression of PEMT2 inhibits the phosphorylation and translocation of PLC gamma 1].

OBJECTIVES: To explore the mechanism of CBRH-7919 cell proliferation inhibition by transfecting phosphatidylethanolamine N-methyltransferase 2 gene (PEMT2). METHODS: The effects of PEMT2 transfection on phosphorylation and translocation from cytosol to plasma membrane of PLC gamma 1 in cells were studied using SDS-PAGE and Western blot techniques. The phosphorylation and activity of c-Met were determined. RESULTS: After transfection of pemt2, the PLC gamma 1 and phosphorylated PLC gamma 1 conjugated with plasma membrane were decreased by 45% and 27% of that of control cells respectively, and the phosphorylated c-Met was decreased to 32% of that of control cells. CONCLUSION: Transfection of phosphatidylethanolamine N-methyltransferase 2 gene can inhibit the phosphorylation and translocation from cytosol to plasma membrane of PLC gamma 1 in cells. At the same time, the autophosphorylation of c-Met was decreased, which suggests that transfection of phosphatidylethanolamine N-methyltransferase 2 gene can downregulate the c-Met/PLC gamma 1 signaling pathway in CBRH-7919 cells.