Panoply™ Human CDH6 Over-expressing Stable Cell Line

N6-methyladenosine (m6A) is involved in biliary atresia (BA), but the underlying mechanisms remain unclear. Here, researchers investigated the involvement of m6A modification and its role in epithelial-to-mesenchymal transition (EMT) in BA cholangiocytes. Single-cell RNA sequencing data revealed significant enrichment of EMT- and fibrosis-related pathways in BA cholangiocytes. Immunofluorescence confirmed upregulation of EMT and fibrosis markers in BA cholangiocytes. M6A-regulated genes were downregulated in BA cholangiocytes, with METTL3 identified as a key regulator. CDH6 was identified as a candidate gene regulated by m6A modification. Functional experiments demonstrated that METTL3-mediated hypomethylation stabilized CDH6 transcripts through YTHDF2. METTL3 downregulation enhanced cholangiocyte migration and EMT progression. RNA-seq and in vitro experiments demonstrated that CDH6 upregulation promoted cholangiocyte proliferation and migration, thereby promoting EMT. Thus, these studies highlight the role of m6A methylation in regulating CDH6 expression and its contribution to EMT progression in cholangiocarcinoma, providing new evidence on potential mechanisms underlying BA.

METTL3 overexpression inhibited wound healing, while METTL3 knockdown enhanced wound healing (Figure 1A). To further investigate the role of CDH6 in m6A-enhanced cell migration, researchers constructed CDH6-disturbed human intrahepatic biliary epithelial cells (HIBECs) and verified its expression (Figure 1B). CCK-8 assays demonstrated that CDH6 overexpression promoted cell proliferation (Figure 1C). Furthermore, Transwell and wound healing assays confirmed enhanced cell migration in CDH6-overexpressing cells (Figures 1D-F). To explore the downstream pathways regulated by CDH6, researchers performed RNA sequencing on CDH6-overexpressing cells and control HIBECs, identifying 233 upregulated genes (Figure 1G). GO and KEGG enrichment analysis revealed that these genes were significantly enriched in EMT-related pathways, including regulation of EMT, epithelial cell migration and proliferation, cell adhesion, and TGF-β signaling. Classical hepatocellular carcinoma-associated pathways (such as Hedgehog and Notch signaling) and epithelial differentiation pathways were also significantly enriched. Furthermore, DEGs in CDH6-overexpressing HIBECs were involved in pathways associated with liver fibrosis and inflammation (Figure 1H). Together, these findings suggest that CDH6 upregulation contributes to EMT activation, cell migration, and BA-related signaling pathways, further implicating it in hepatic fibrosis and inflammation.

Figure 1. Upregulation of CDH6 devoted to cell migration and EMT pathway. (Meng L, et al., 2025)