Abstract
Hepatocellular carcinoma (HCC) requires novel therapeutic targets, and oncofetal genes offer a promising avenue. Although the adaptor protein AP2M1 is a known prognostic factor in HCC, its functional role in promoting cancer stemness remains unclear. This study investigates this role using an integrative in silico, in vitro, and in vivo approach. We combined TCGA database analysis with experiments using AP2M1 knockdown in HCC cell lines (HepG2, Hep3B) and morpholino knockdown of its homolog, ap2m1a, in zebrafish embryos. In silico analysis showed elevated AP2M1 in HCC, correlating positively with stemness markers. In vitro, AP2M1 knockdown suppressed proliferation, migration, invasion, and the expression of key stemness genes like DLK1 and NANOG. These findings were recapitulated in vivo, where ap2m1a knockdown in zebrafish impaired hepatocyte development and downregulated stemness markers. Mechanistically, AP2M1 depletion consistently suppressed the Wnt signaling pathway, evidenced by reduced AXIN2 expression across all models. Collectively, this study establishes AP2M1 as a key regulator of stemness in HCC, operating through the Wnt signaling pathway. Our findings validate AP2M1 as a promising biomarker and potential therapeutic target for HCC.