A Molecular and Computational study on the effects of 5-Fluorouracil , and Dicyclomine on AGS Cells: Regulation of P53 and SNAIL2 Pathways

Abstract

Background: 5-Fluorouracil (5-FU) is commonly used to inhibit gastric cancer (GC) cell growth, but its clinical effectiveness is frequently limited by chemoresistance. Dicyclomine (DIC), an anticholinergic agent, has been suggested to enhance the therapeutic effect of 5-FU in cancer treatment. Objectives: This study aimed to investigate the effects of 5-FU, DIC, and their combination on AGS cell viability and the expression of SNAIL2 and P53. Methods and material: AGS cells were cultured in Dulbecco's Modified Eagle Medium/F12 supplemented with 10% fetal bovine serum (FBS). Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The mRNA expression levels of P53 and SNAIL2 were analyzed by quantitative real-time PCR (qRT-PCR). Drug synergism was evaluated using the CompuSyn software. Network pharmacology was employed to investigate the potential relationship between DIC and the expression of P53 and SNAIL2 genes. Results: The IC₅₀values for 5-FU and DIC were 4 µg/mL and 800 µg/mL, respectively. In AGS cells, all treatments significantly upregulated P53 expression (5-FU: p<0.01; DIC: p<0.001; Combination: p<0.0001). While 5-FU (p<0.01) or DIC (p<0.0001) alone increased SNAIL2 expression, their combination significantly reduced it (p<0.05). CompuSyn software confirmed a synergistic interaction. KEGG pathway analysis revealed 38 enriched pathways, with the adherens junction pathway ,highlighted due to its potential role in SRC protein-mediated downregulation of SNAIL gene expression. Conclusion: These findings indicate that the combination of 5-FU and DIC synergistically exerts significant anticancer effects by upregulating P53 and downregulating SNAIL2 expression, representing a promising strategy for optimizing GC management.

Keywords: Gastric neoplasms, P53, SNAIL2, Fluorouracil, Dicyclomine