4-Acetylantrocamol LT3 Suppresses Colorectal Cancer Growth and Metastasis via PI3K/AKT and MAPK Pathway Modulation

Background: Colorectal cancer (CRC) remains one of the leading causes of cancer-related mortality worldwide. Despite advances in targeted therapies, drug resistance and limited efficacy in KRAS-mutant CRC continue to present clinical challenges. Antrodia cinnamomea, a medicinal fungus, has demonstrated anti-tumor properties, yet the mechanisms of its triterpenoid compound 4-acetylantrocamol LT3 (LT4) remain unclear. Methods: We investigated LT4’s effects in HCT116 CRC cells using cell viability, colony formation, and migration assays. Western blotting was employed to examine key signaling pathways. Transcriptome profiling via RNA sequencing was followed by KEGG/GO enrichment and protein–protein interaction (PPI) network analyses using STRING, CytoHubba, and MCODE. Molecular docking with PI3Kγ (PDB: 1E7U) was conducted to evaluate LT4’s binding affinity. Results: LT4 significantly inhibited HCT116 cell proliferation and migration, induced a mesenchymal-to-epithelial transition, suppressed PI3K/AKT/mTOR and ERK signaling, and activated GSK3β/FoxO and p38/p21 axes. It also reduced COX-2 and anti-apoptotic proteins (Bcl-2, Bcl-XL), while impairing mitochondrial stability (COX IV). Transcriptomic analysis identified the PI3K-Akt pathway as the most significantly enriched signaling cascade. Network topology analyses highlighted SLC3A2, CCND1, PSAT1, and CHAC1 as central nodes, linking LT4’s effects to nutrient signaling, redox homeostasis, and serine metabolism. Molecular docking confirmed that LT4 stably occupied the ATP-binding pocket of PI3Kγ with a binding energy comparable to KWT and a conformation similar to Antroquinonol. Conclusion: This study is the first to comprehensively demonstrate LT4’s multi-target anti-CRC effects, highlighting its potential as a therapeutic agent, especially in KRAS-mutant CRC.