Objective <p>This study aimed to investigate the function and role of PPP2CA in the colon by analyzing the phenotype of colon-specific PPP2CA-knockout mice. PPP2CA is the gene encoding the catalytic subunit of serine/threonine protein phosphatases 2A (PP2A).</p> Methods <p>A conditional knockout mouse model was established using the Cre/loxP system. Physiological changes in mice were recorded after tamoxifen-induced knockout. Phenotypic differences between wild-type mice and conditional knockout mice were observed through anatomical, molecular biological, label-free quantitative proteomic, and histological methods.</p> Results <p>There were no significant differences in body weight and defecation between the knockout mice and wild-type mice. However, at the pathological level, the knockout group exhibited a higher abundance of goblet cells and increased proliferation of intestinal villus cells. Proteomic analysis indicated alterations in the expression of key proteins involved in processes such as metabolic reprogramming and immune response.</p> Conclusion <p>The deletion of PPP2CA in the colon did not cause intestinal dysfunction; instead, it may enhance intestinal barrier function by promoting the proliferation of cells related to the mucosal barrier.</p>