Network pharmacology integrated with multi-omics demonstrates that Wumei Wan modulates the progression of colorectal cancer by regulating the focal adhesion-YAP signaling axis.

[ETHNOPHARMACOLOGICAL RELEVANCE] Colorectal cancer(CRC) is one of the most common malignant tumors of the digestive system, and Traditional Chinese Medicine has shown good efficacy in its treatment and prognosis. Wumei Wan(WMW) is widely used in the treatment of digestive system and tumor diseases, with effects such as anti-diarrheal, anti-dysentery, and pain relief. Modern pharmacological studies suggest that its active ingredients have anti-inflammatory, antioxidant, and anti-tumor properties. However, there is still a lack of systematic research on the role of WMW in the prevention and treatment of CRC and its molecular mechanisms.

[AIM OF THIS STUDY] The aim of this study is to clarify the key active components and primary mechanisms by which WMW inhibits the initiation and progression of CRC, utilizing a multi-faceted approach including chemical composition analysis, network pharmacology, multi-omics studies, molecular biology validation, and molecular simulations.

[METHODS] UHPLC-HRMS/MS was used to identify the chemical components in WMW decoction and post-administration serum, and network pharmacology analysis was combined to predict potential targets and signaling pathways. Key differential pathways were selected through combined transcriptomics and proteomics analysis, and the expression patterns of the FAK/Src/YAP axis in CRC were verified using the GEO database. An azoxymethane/dextran sulfate sodium (AOM/DSS)-induced CRC mouse model was constructed to assess the effects of WMW on survival rate, tumor burden, histological changes in the colon, and the expression of related proteins. The expression of downstream genes CTGF and Axin2 was detected via qRT-PCR, and the regulatory effects of WMW-containing serum on FAK/Src/YAP signaling were examined in the HCT116 cells model. Finally, molecular docking and molecular dynamics simulations were performed.

[RESULTS] UHPLC-HRMS/MS analysis identified 15 circulating components of WMW following administration. Network pharmacology analysis revealed 241 overlapping targets between WMW-related targets and CRC-associated genes, which were enriched in pathways related to inflammation regulation, cell proliferation, and focal adhesions, with FAK/Src/YAP-related signaling features emerging as a central network module. In the AOM/DSS-induced CRC model, WMW treatment was associated with changes in body weight, tumor burden, and colonic mucosal structure, along with reduced tissue expression levels of VEGF and p53. Multi-omics analyses showed coordinated alterations in pathways related to inflammation, proliferation, and extracellular matrix (ECM) remodeling following WMW intervention. Analysis of GEO datasets confirmed elevated expression of PTK2, Src, and YAP1 in CRC tissues, supporting aberrant focal adhesion-YAP-associated signaling features in clinical CRC. In vivo and in vitro experiments showed that WMW intervention was associated with reduced phosphorylation levels of FAK, Src, and YAP, decreased stress fiber formation, attenuated YAP nuclear localization, and lower expression of CTGF and PCNA. Molecular docking and molecular dynamics simulations indicated that several circulating components could stably interact with the FAK kinase domain, with auraptene exhibiting relatively favorable binding characteristics.

[CONCLUSION] These findings suggest that WMW intervention is associated with coordinated alterations in inflammation-related signaling, cell proliferation, and extracellular matrix remodeling in CRC, with focal adhesion-YAP-related mechanotransduction representing a potential associated feature. This study provides integrated experimental evidence supporting further investigation of WMW and traditional Chinese medicine-based strategies targeting focal adhesion-associated signaling in CRC research.