<p>Despite significant advances in colorectal cancer (CRC) diagnosis and treatment, drug therapy of CRC patients is still confronted with considerable challenges. Carbohydrate response element-binding protein (ChREBP), a glucose-responsive transcription factor regulating glycolysis and de novo lipogenesis, shows elevated expression in human CRC tissues and correlates with poor disease-free survival and overall survival. However, the in vivo role and mechanism of ChREBP in colorectal carcinogenesis remain unclear. We used ChREBP knockout mice, which were intraperitoneally injected with azoxymethane (AOM) followed by dextran sulfate sodium (DSS) in drinking water. In the AOM/DSS-induced colorectal cancer model, carcinogenesis was reduced in ChREBP null mice. In the initial phases of colorectal carcinogenesis, ChREBP deficiency was associated with diminished epithelial cell proliferation and a lower number of aberrant crypt foci, but it had no impact on DNA damage or the severity of colitis. The key transcription factor β-catenin and Wnt target gene expression were both decreased in the colons of ChREBP null mice and in ChREBP-knockdown Caco-2 colorectal cancer cells. In vitro studies demonstrated that ChREBP overexpression promoted β-catenin accumulation, nuclear translocation, and transcriptional activity by interacting with β-catenin, while ChREBP knockdown produced the opposite effects. These findings establish a novel mechanism whereby ChREBP drives CRC progression through Wnt/β-catenin pathway activation, positioning it as both a potential therapeutic target and prognostic biomarker for CRC.</p><p></p>