Based on the Wnt/β-catenin pathway, to investigate the anti-tumor effect of Gegen Qinlian Decoction (GQD) on colorectal cancer and its effect on nuclear translocation of protein regulator of cytokinesis 1 (PRC1). BALB/c nude mice were subcutaneously inoculated with the CT26 colorectal cancer cell line and divided into GQD low(L-GQD), medium(M-GQD), and high(H-GQD) dose groups, with the model group serving as the control. The mice were administered the drug daily for 25 days, and the growth of the tumors was recorded. Following the conclusion of the previous administration, the tumor was surgically excised and subjected to subsequent observation. The proliferation and apoptosis of the tumor were assessed using hematoxylin-eosin staining(HE), immunohistochemistry (IHC), and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining techniques, while the Wnt/β-catenin pathway was examined through qRT-PCR and western blot (WB) analysis. Furthermore, alterations in the expression of catenin signals and downstream factors were investigated. In addition, WB was used to detect the effects of GQD on the phosphorylation level of PRC1 and its subcellular localization. Compared with the model group, GQD inhibited the growth and increased the apoptosis level of colorectal tumors in vivo in a dose-dependent manner. IHC results show that GQD down-regulated the expressions of proliferation-related proteins cyclind1, marker of proliferation Ki-67 (Ki67), and platelet endothelial cell adhesion molecule-1 (CD31) (P < 0.05), and promoted the expressions of approbation-related proteins Caspase 3 and Caspase 9(P < 0.05). GQD is further found to inhibit the activation of Wnt/β-catenin signaling, which may be related to the reduction of PRC1 phosphorylation level and the alteration of its nuclear retention ratio. This inhibition of Wnt/β-catenin signaling by GQD is closely associated with the suppression of colorectal tumor proliferation and the promotion of apoptosis. Moreover, GQD may involve in mediating nuclear translocation of PRC1.