Baicalin suppresses colorectal cancer proliferation and induces M1 polarization of tumor-associated macrophages by promoting proteasomal degradation of HK2

Colorectal cancer (CRC) ranks as the third most diagnosed cancer and the second leading cause of cancer-related mortality globally. Hexokinase II, a key rate-limiting enzyme in tumor glycolysis, is an important therapeutic target. In this study, we report that baicalin, a flavonoid derived from Scutellaria baicalensis, acts as a HK2 inhibitor and exerts anti–colorectal cancer activity. In vitro, baicalin markedly suppressed colorectal cancer cell proliferation and colony formation. Molecular docking, molecular dynamics simulations, DARTS, and CETSA suggest an association between baicalin and HK2, while MG132 rescue and HK2 immunoprecipitation indicate that baicalin promotes ubiquitination-associated proteasomal degradation of HK2. Mechanistically, baicalin inhibits HK2, reduces glycolysis, and causes mitochondrial damage, thereby activating the cGAS/STING innate immune signaling pathway and increasing IFN-β production. IFN-β contributes to reshaping the tumor immune microenvironment. In an MC38 syngeneic tumor model, baicalin significantly inhibited tumor growth, reduced HK2 protein levels, activated the cGAS/STING pathway, and promoted a shift in tumor-associated macrophages toward an M1-like polarization state. Collectively, this study uncovers a novel strategy for targeting HK2 to regulate both tumor cell metabolism and the immune microenvironment, providing a potential therapeutic approach for colorectal cancer.