Deregulated expression of N-acetylgalactosaminyltransferases (GALNTs), which responsible for initiating step of mucin-type O-glycosylation, could produce abnormal truncated O-glycans and thereby exerting pivotal functions during malignant transformation. GALNT4 is one of the few isoforms preferring to catalyze partial GalNAc-glycosylated substrates and modify the sites not utilized by other known GALNTs. This study aims to evaluate the impact of GALNT4 expression on malignant transformation of hepatocellular carcinoma (HCC). Immunohistochemistry and in situ hybridization analysis were performed to assess GALNT4 and miR-9 level in clinical specimens, respectively. GALNT4 expression is markedly repressed in primary HCC tissues and reduced expression of GALNT4 is significantly associated with adverse survival of patients with HCC. Functional investigations demonstrate that repressed GALNT4 could promote migration, invasion, anoikis resistance and stemness of HCC cells in vitro as well as tumor growth in vivo. The wild-type GALNT4 could modify O-linked glycosylation on EGFR and thus modulate the activity of EGFR. Luciferase activity assay further identified microRNA-9 (miR-9) as the crucial specific arbitrator for GALNT4 expression in HCC cells. Furthermore, restoring GALNT4 expression attenuates miR-9 mediated oncogenic functions. Kaplan-Meier survival analysis indicates the miR-9/GALNT4 expression signature yield promising prognostic significance to refine the risk stratification of patients with HCC. In conclusions, this study establishes miR-9/GALNT4 axis as a potential adverse prognostic factor and therapeutic target for HCC patients.
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