Exploring the pharmacological mechanism of Xianlian Jiedu decoction in the treatment of colorectal Cancer based on network pharmacology, molecular docking and experimental validation

Abstract

BackgroundXianlian Jiedu Decoction (XLJDD) is a clinical effective prescription for the treatment of colorectal cancer (CRC) for several decades. However, the effective mechanism and targets of action of XLJDD are still unclear. This study aims to discover the pharmacological mechanism of XLJDD by employing a combination of network pharmacology, molecular docking and in vitro and in vivo experimental validation. MethodsMultiple databases, including TCMSP, Swiss Target Prediction, Drugbank, Geencards, Malacards, Therapeutic Target Database (TTD) and Disgenet, were searched and identified the targets of ingredients from XLJDD and the targets associated with CRC. The common targets of ingredients and disease were analyzed, and the visualization network of protein-protein interaction (PPI) were constructed using STRING database and to identified the core targets based on Cytoscape software. Metascape database was employed for GO and KEGG enrichment analysis of common targets. The active ingredients of XLJDD and core targets were verified through molecular docking test. Subsequently, in vitro and in vivo experiments were performed to validate the analysis results using CT-26 and HCT116 cell lines and AOM/DSS mice model. ResultsA total of 36 active components and 122 targets were identified for XLJDD, while 1728 targets associated with CRC were obtained. Among them, there were 71 common targets between ingredients and disease. And 19 core targets (AKT1, TNF, PTGS2, CASP3, EGFR, ESR1, MMP9, IL1B, CCND1, PPARG, ERBB2, SIRT1, MMP2, BCL2L1, MAPK1, PPARA, PGR, AR and SERPINE1.) were finally screened from PPI network. GO analysis enriched a total of 99 entries, with 74 related to biological processes, 20 to molecular functions, and 5 to cellular components. The KEGG enrichment pathway analysis demonstrated that the biological discrepancy was mainly focused on the tumorigenesis-, inflammation-, and mechanism-related pathways. Molecular docking presented the great bonding ability of 19 key targets with 17 active constituents. Experiments based on CT-26 and HCT116 cells verified that XLJDD could inhibit the proliferation of colorectal cancer cells. In addition, XLJDD inhibited tumor progression in mice. Finally, the results of molecular biology experiments showed that XLJDD could activate the colorectal cancer-related PTEN/AKT/FoXO1 pathway, and up-regulate the expression levels of key genes such as AKT1, TNF, PTGS2 and down-regulate the expression levels of CASP3. ConclusionsThis study revealed comprehensively the targets and mechanism for the efficacy of XLJDD in treatment of CRC. Furthermore, the results were achieved through in vitro experiment of CT-26 and HCT116 cell lines, and in vivo experiment of AOM/DSS mice.