文献类型：期刊文献

英文题名：Investigating the Multi-Target Mechanisms of Calycosin-7-O-β-D-glucoside Against Colorectal Cancer via Integrated Network Pharmacology, Molecular Docking, and In Vitro Experiments

作者：Li, Zhaohuan[1,2];Xu, Jie[3];Guo, Jianjin[4];Zhang, Xin[1];Zhao, Jingwen[5];Li, Peihan[6];Gui, Mingbin[7];Cheng, Xin[8];Gao, Feng[1,2]

第一作者：Li, Zhaohuan

通信作者：Gao, F[1];Gao, F[2];Guo, JJ[3];Cheng, X[4]

机构：[1]Gansu Univ Chinese Med, Sch Clin Chinese Med, Lanzhou 730000, Peoples R China;[2]Gansu Prov Cent Hosp, Dept Colorectal & Anal Surg, Lanzhou 730050, Peoples R China;[3]First Hosp Jilin Univ, Dept Hepatobiliary & Pancreat Surg 1, Changchun 130021, Peoples R China;[4]Lanzhou Univ, Coll Chem & Chem Engn, State Key Lab Nat Prod Chem, Lanzhou 730000, Peoples R China;[5]Xian Hosp Tradit Chinese Med, Dept Colorectal & Anal Surg, Xian 710021, Peoples R China;[6]Qinghai Prov Hosp Tradit Chinese Med, Dept Anorectal Dis 1, Xining 810000, Peoples R China;[7]940th Hosp Joint Logist Support Force Chinese Peop, Dept Colorectal & Anal Surg, Lanzhou 730050, Peoples R China;[8]Shaanxi Prov Peoples Hosp, Dept Gen Surg 2, Xian 710068, Peoples R China

第一机构：甘肃中医药大学

通信机构：[1]corresponding author), Gansu Univ Chinese Med, Sch Clin Chinese Med, Lanzhou 730000, Peoples R China;[2]corresponding author), Gansu Prov Cent Hosp, Dept Colorectal & Anal Surg, Lanzhou 730050, Peoples R China;[3]corresponding author), Lanzhou Univ, Coll Chem & Chem Engn, State Key Lab Nat Prod Chem, Lanzhou 730000, Peoples R China;[4]corresponding author), Shaanxi Prov Peoples Hosp, Dept Gen Surg 2, Xian 710068, Peoples R China.|[10735]甘肃中医药大学;

年份：2026

卷号：22

期号：1

外文期刊名：JOURNAL OF PHARMACEUTICAL INNOVATION

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001883832800001)】；

基金：This work was supported by the Gansu Provincial Natural Science Foundation (Key Research and Development Program Project, Grant 20YF8FA098; Basic Research Program Project, Grant 23JRRA539), the Gansu Provincial Youth Science and Technology Fund (Grant 25JRRA429), and the Lanzhou Municipal Science and Technology Bureau Project (Grant 2024-9-163).

语种：英文

外文关键词：Calycosin-7-O-beta-D-glucoside; Colorectal cancer; PI3K-Akt signaling pathway; Network pharmacology; Molecular dynamics simulation; Multi-target mechanisms

摘要：Purpose Colorectal cancer (CRC) is a heterogeneous and drug-resistant malignancy that urgently requires multi-target therapeutic agents. This study aimed to systematically elucidate the multi-target molecular mechanisms of calycosin-7-O-beta-D-glucoside (CG), a bioactive flavonoid from Astragali radix, against CRC by integrating computational prediction, dynamic simulation, clinical bioinformatics validation, and in vitro experimental verification.Methods Network pharmacology was employed to predict CG targets and CRC-associated genes, with overlapping targets subjected to PPI network construction, GO/KEGG enrichment analyses, and core target screening. Molecular docking and 100-ns molecular dynamics (MD) simulation were performed to evaluate binding affinity and complex stability. Multi-database bioinformatics analyses (HPA, GEPIA, TIMER) validated clinical relevance, prognostic value, and immune infiltration correlations of core targets. In vitro assays, including CCK-8, TUNEL, EdU, flow cytometry, colony formation, wound healing, Transwell invasion, tumorsphere formation, and Western blotting, were conducted in HCT116 and HT29 cells to confirm CG's antitumor effects and mechanistic regulation.Results Network pharmacology identified 99 overlapping targets, with ten core targets and significant enrichment in the PI3K-Akt signaling pathway. Molecular docking revealed strong binding affinities, and MD simulation confirmed long-term stability of the CG-HSP90AA1 complex. Multi-database analyses showed core target overexpression in CRC tissues, association with poor prognosis, and correlations with CD8(+) T cells, Tregs, MDSCs, and neutrophils. In vitro, CG concentration-dependently suppressed viability, induced apoptosis, inhibited proliferation, migration, invasion, and tumorsphere formation. Western blotting confirmed that CG reduced p-PI3K/PI3K and p-Akt/Akt ratios, decreased Bcl-2/Bax, and activated cleaved caspase-3, triggering the mitochondrial apoptotic pathway.Conclusion This study is the first to demonstrate that CG exerts anti-CRC effects through multi-target binding to HSP90AA1, TNF, and PTGS2, coordinately suppressing the PI3K-Akt pathway and downstream apoptotic cascades, thereby inhibiting malignant phenotypes and cancer stem cell-like properties. These findings establish a comprehensive "computational prediction -> dynamic simulation -> clinical association -> experimental validation" paradigm and provide experimental evidence supporting CG as a promising adjuvant therapeutic candidate for CRC.