Berberine Inhibited Tumor Growth Through Suppressing the WDR54/β‐catenin Signal Pathway in Colorectal Cancer

ABSTRACT

Berberine (BBR), a natural alkaloid, exhibits potent anti‐tumor activity in colorectal cancer (CRC), but its mechanisms are not fully elucidated. Analysis of GEO datasets (GSE216908 and GSE184414) identified WDR54 as a key target of BBR treatment. In vitro, BBR treatment of CRC cell lines (HT‐29 and SW480) markedly inhibited cell proliferation, DNA synthesis, and colony formation. Notably, BBR triggered G1‐phase cell cycle arrest and promoted apoptosis, as validated by flow cytometry and molecular marker analysis. WDR54 overexpression partially rescued these anti‐tumor effects, confirming its functional importance. Mechanistically, cellular thermal shift assays (CETSA) and mRNA stability assays revealed that BBR suppresses WDR54 expression via indirect transcriptional inhibition rather than direct protein binding or altered mRNA stability. Dual‐luciferase reporter assays further confirmed that BBR dose‐dependently inhibits WDR54 promoter activity. In vivo, an AOM/DSS‐induced CRC mouse model showed that BBR reduced tumor number and size, alleviated pathological progression, and suppressed the WDR54/β‐catenin axis. These findings suggest that BBR exerts significant anti‐tumor effects in CRC by downregulating WDR54 and inhibiting Wnt/β‐catenin signaling, highlighting WDR54 as a potential therapeutic target and BBR as a promising treatment for CRC.