Tabriz University of Medical Sciences Pharmaceutical Sciences 1735-403X 32 2 2026 04 30 A Molecular and Computational study on the effects of 5-Fluorouracil, and Dicyclomine on AGS Cells: Regulation of P53 and SNAIL2 Pathways 291 301 10.34172/PS.026.43173 EN Fariba Ghodrati https://orcid.org/0000-0002-4393-9734 Mahdi Bagheri Seyedeh Yasaman Kolki Parham Motamedi Elham Sadeghi Hossein Ebrahimi Dhifaf Saleem Kareem Maliki Hadis Musavi https://orcid.org/0009-0000-2523-4653 Journal Article 10.34172/PS.026.43173 2025 09 11 2025 12 15 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: 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 IC₅₀ values 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.