O-GlcNAcylation of progranulin promotes hepatocellular carcinoma proliferation

Progranulin (PGRN) is overexpressed and implicated in hepatocellular carcinoma (HCC) development; however, its post-translational modifications and regulatory mechanisms in HCC remain largely unexplored. Here, the expression levels of PGRN, OGT, and O-GlcNAcylation were found to be elevated in both HCC samples and cell lines. LC-MS/MS analysis and immunoprecipitation revealed that PGRN underwent O-linked N-acetylglucosamine (O-GlcNAc) modification at threonine 272 (Thr272). Co-immunoprecipitation and confocal microscopy confirmed the interaction and colocalization of O-GlcNAc transferase (OGT) with PGRN. Reducing O-GlcNAcylation increased the ubiquitination of PGRN, while increasing O-GlcNAcylation inhibited ubiquitination and elevated PGRN stability, as measured by cycloheximide (CHX) chase experiments. This regulation of PGRN stability was directly linked to its expression levels. Moreover, mutation at the primary O-GlcNAc site Thr272 inhibited the activity of the PI3K/AKT/mTOR signaling pathway and suppressed HCC cell proliferation. Together, our findings indicate that O-GlcNAcylation at Thr272 is essential for PGRN-driven HCC cell proliferation. [Display omitted] •PGRN is O-GlcNAcylated at Thr272, enhancing its stability in HCC cells.•Thr272 O-GlcNAcylation inhibits PGRN ubiquitination and promotes its expression.•O-GlcNAcylation at Thr272 activates the PI3K/AKT/mTOR pathway to drive HCC proliferation.•Mutation of Thr272 reduces HCC cell viability and colony formation.•O-GlcNAcylation of PGRN is a potential therapeutic target in HCC.