Hesperetin as a potential therapeutic agent for colorectal cancer: Targeting PI3K/AKT and EMT in cell lines through network pharmacology-guided strategies

Hesperetin is a naturally occurring flavonoid compound abundantly present in citrus fruit peels and Traditional Chinese Medicinal (TCM). It exhibits diverse pharmacological properties and represents a promising multi-target candidate for anticancer therapy. This study systematically investigated the molecular mechanisms underlying the anti-colorectal cancer (CRC) effects of hesperetin by integrating network pharmacology, molecular docking, and experimental validation. Network pharmacology analysis identified 42 core targets of hesperetin in CRC, with molecular docking confirming strong binding affinities (binding energy < −7 kcal/mol) between hesperetin and key proteins, including Epidermal Growth Factor Receptor (EGFR), Threonine Kinase 1 (AKT1), Proto-oncogene Tyrosine-protein Kinase Src (SRC), Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha (PIK3CA), and Matrix Metalloproteinase-9 (MMP9). In vitro experiments demonstrated that hesperetin dose-dependently inhibited the proliferation, migration, and invasion of HCT116 and Lovo cells. Mechanistically, hesperetin reversed epithelial-mesenchymal transition (EMT) by upregulating E-cadherin and downregulating N-cadherin, Vimentin, and Snail at both protein and mRNA levels. Western blot analysis revealed that hesperetin suppressed PI3K/AKT pathway activation by reducing phosphorylation of PI3K (Tyr458) and AKT (Ser473). Clinical data further validated the therapeutic relevance of these targets, showing that high expression of EGFR, AKT1, PIK3CA, and MMP9 correlated with poor prognosis in CRC patients. Collectively, these findings establish hesperetin as a promising multi-target nutritional supplement agent against CRC, demonstrating dual modulation of both PI3K/AKT signaling and EMT progression.