Ginkgolide B Inhibits Colorectal Cancer Growth via Regulation of the PI3K/AKT/MAPK Pathways: A Comprehensive Analysis Using Network Pharmacology, Molecular Docking, and Verification in Cellular and Animal Models

Background: Ginkgolide B (GB), a bioactive constituent isolated from Ginkgo biloba, has shown antitumor activity in several malignancies. However, the molecular mechanisms underlying GB action against colorectal cancer (CRC) remain incompletely understood. Thus, this study aimed to systematically investigate the potential molecular mechanisms and therapeutic effects of GB on CRC using an integrated approach. Methods: Potential therapeutic targets of GB against CRC were identified by integrating GB-related and CRC-associated genes. Protein–protein interaction (PPI) networks and functional enrichment analysis (FEA) identified key targets and pathways. Interactions between GB and hub proteins were assessed by docking. The antitumor effects of GB were evaluated in CRC cells using assays for cell proliferation, colony formation, and apoptosis, as well as Western blotting. These data were validated in an MC38 syngeneic subcutaneous tumor model. Results: A total of 288 target proteins were shared between the 522 predicted GB targets and the 4199 CRC-associated targets. FEA indicated that these targets were primarily associated with the phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT) and mitogen-activated protein kinase (MAPK) axes. Docking predicted favorable binding of GB to several hub proteins, with AKT1 showing the strongest predicted affinity. Experimental validation demonstrated that GB inhibited CRC cell proliferation and colony formation, promoted apoptosis, decreased B-cell lymphoma 2 (BCL-2), increased BCL-2-associated X protein (BAX) and Cleaved Caspase-3, and reduced the phosphorylation of AKT and extracellular signal-regulated kinase (ERK). Notably, GB significantly suppressed tumor growth and reduced Ki-67 expression in the MC38 syngeneic subcutaneous tumor model. Conclusions: GB exhibits antitumor activity against CRC by inhibiting proliferation, promoting apoptosis, and inhibiting the PI3K/AKT and MAPK pathways. The study provides a mechanistic basis for the anticancer effects of GB and supports further investigation of its potential as a therapeutic candidate for CRC.