Yan Sicheng, Hu Xiaomeng, Wu Yan, Ye Wangfang, Zhu Yuehong, He Yuxuan, Zhan Fuyuan, Wu Wei, Ma Zhihong
Huzhou Key Laboratory of Molecular Medicine, Huzhou Central Hospital, Fifth School of Clinical Medicine of Zhejiang Chinese Medical University, Huzhou, Zhejiang, 313000, People's Republic of China.
School of Basic Medicine College, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310000, People's Republic of China.
Drug Des Devel Ther. 2025 May 20;19:4105-4122. doi: 10.2147/DDDT.S474421. eCollection 2025.
5-Fluorouracil (5-FU) is a mainstream drug used in chemotherapy and chemoradiotherapy regimens for the clinical treatment of malignancies, such as gastric cancer (GC), colorectal cancer, and breast cancer. However, the molecular mechanism of action of 5-FU in GC has not yet been studied using a network pharmacology approach.
The mechanism of action of 5-FU in GC was determined using a network pharmacology technique, and our findings were confirmed by various computational approaches and experimental tests using the GeneCards database, ChEMBL database, STRING database, molecular docking, molecular dynamics simulation, DAVID, GEPIA, Kaplan‒Meier Plotter, CCK-8 assays, colony formation experiments, cell proliferative assay, apoptosis assays, wound-healing assays, Real-time PCR and Western blot tests.
A total of 21 shared and 13 potential targets were identified using PPI network analysis. Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses indicated that the PI3K/AKT signaling pathway may be a significant pathway. Combined with molecular docking and database verification, F10, NR3C1, DHFR, CA2, BCHE, ACHE, and ITGA4 were identified as candidate core genes. Moreover, the experimental results illustrated that ITGA4 induces 5-FU resistance by up-regulating PI3K/AKT signaling.
Network pharmacology is a feasible scientific research strategy for revealing the multitarget-multipathway role of 5-FU in the treatment of GC and provides ITGA4-based new ideas and therapeutic strategy to overcome 5-FU resistance for GC treatment.
5-氟尿嘧啶（5-FU）是用于恶性肿瘤临床治疗的化疗和放化疗方案中的主流药物，如胃癌（GC）、结直肠癌和乳腺癌。然而，尚未采用网络药理学方法研究5-FU在GC中的分子作用机制。
采用网络药理学技术确定5-FU在GC中的作用机制，并通过各种计算方法和实验测试进行验证，这些方法和测试使用了基因卡数据库、ChEMBL数据库、STRING数据库、分子对接、分子动力学模拟、DAVID、GEPIA、Kaplan-Meier Plotter、CCK-8测定、集落形成实验、细胞增殖测定、凋亡测定、伤口愈合测定、实时PCR和蛋白质免疫印迹测试。
通过蛋白质-蛋白质相互作用（PPI）网络分析共鉴定出21个共享靶点和13个潜在靶点。京都基因与基因组百科全书（KEGG）富集分析表明，PI3K/AKT信号通路可能是一条重要通路。结合分子对接和数据库验证，确定F10、NR3C1、二氢叶酸还原酶（DHFR）、碳酸酐酶2（CA2）、丁酰胆碱酯酶（BCHE）、乙酰胆碱酯酶（ACHE）和整合素α4（ITGA4）为候选核心基因。此外，实验结果表明，ITGA4通过上调PI3K/AKT信号诱导5-FU耐药。
网络药理学是揭示5-FU在GC治疗中多靶点-多途径作用的可行科研策略，并为克服GC治疗中的5-FU耐药性提供了基于ITGA4的新思路和治疗策略。
