OBJECTIVE To explore the inhibitory effect of Huangqi-Ezhu combination on the progression of colon cancer by regulating P-selectin-mediated adhesion and migration.

METHODS The GSE126095 dataset from the GEO database was used to conduct differential expression analysis, KEGG pathway enrichment analysis, and P-selectin correlation analysis. In cell experiments, CT26 cells were transfected with P-selectin overexpression plasmid and intervened with 15% Huangqi-Ezhu medicated serum. CCK-8 assay, FDA staining, Transwell assay and wound healing assay were performed to detect cell proliferation, adhesion and migration abilities. In animal experiments, an orthotopic colon cancer model was constructed in C57BL/6 mice with MC38 cells. The mice were divided into normal group, model group, low-dose (3 g·kg⁻¹) and high-dose (6 g·kg⁻¹) Huangqi-Ezhu combination groups, with 6 mice in each group. All mice were treated by intragastric administration for 21 days. Western blot, immunohistochemistry and immunofluorescence were applied to detect the expression levels of EMT-related proteins, P-selectin, and adhesion and migration-related proteins in tumor tissues.

RESULTS GEO bioinformatics analysis showed that the differentially expressed genes in colorectal cancer were mainly enriched in Pathways in cancer, and the top 15 differentially expressed genes in this pathway were significantly correlated with P-selectin expression. P-selectin overexpression markedly enhanced the proliferation, adhesion and migration abilities of CT26 cells (P<0.01). Compared with the model group, the tumor volume was significantly decreased in both high-dose and low-dose Huangqi-Ezhu groups (P<0.05, P<0.01). In the high-dose Huangqi-Ezhu group, the vascular expression of P-selectin in tumor tissues was significantly down-regulated (P<0.01), and the abnormal expression of S-Lewisx, PSGL-1, Mucin-1, Integrin and EMT-related proteins was effectively improved (P<0.05, P<0.01).

CONCLUSION Huangqi-Ezhu combination can inhibit tumor growth of orthotopic colon cancer xenografts in mice, down-regulate the expression of vascular P-selectin and adhesion and migration-related molecules in tumor tissues, and ameliorate the abnormal expression of EMT-related proteins.