The oncogenic effect of sulfatase 2 in human hepatocellular carcinoma is mediated in part by glypican 3-dependent Wnt activation.

Publication Type:

Journal Article

Source:

Hepatology (Baltimore, Md.), Volume 52, Issue 5, p.1680-9 (2010)

Keywords:

Animalsdigestive disease, digestive deseases Carcinoma, Hepatocellulardigestive disease, digestive deseases Caspase 3digestive disease, digestive deseases Cell Line, Tumordigestive disease, digestive deseases Enzyme Activationdigestive disease, digestive deseases Flow Cytometrydigestive disease, digestive deseases Gene Expression Regulation, Neoplasticdigestive disease, digestive deseases Genes, Reporterdigestive disease, digestive deseases Glypicansdigestive disease, digestive deseases Humansdigestive disease, digestive deseases Ki-67 Antigendigestive disease, digestive deseases Liver Neoplasmsdigestive disease, digestive deseases Luciferasesdigestive disease, digestive deseases Micedigestive disease, digestive deseases Mice, Nudedigestive disease, digestive deseases Plasmidsdigestive disease, digestive deseases Sulfotransferasesdigestive disease, digestive deseases Transfectiondigestive disease, digestive deseases Wnt Proteinsdigestive disease, digestive deseases Wnt3 Proteindigestive disease, digestive deseases Wnt3A Protein

Abstract:

Heparan sulfate proteoglycans (HSPGs) act as coreceptors or storage sites for growth factors and cytokines such as fibroblast growth factor and Wnts. Glypican 3 (GPC3) is the most highly expressed HSPG in hepatocellular carcinoma (HCC). Sulfatase 2 (SULF2), an enzyme with 6-O-desulfatase activity on HSPGs, is up-regulated in 60% of primary HCCs and is associated with a worse prognosis. We have previously shown that the oncogenic effect of SULF2 in HCC may be mediated in part through up-regulation of GPC3. Here we demonstrate that GPC3 stimulates the Wnt/β-catenin pathway and mediates the oncogenic function of SULF2 in HCC. Wnt signaling in vitro and in vivo was assessed in SULF2-negative Hep3B HCC cells transfected with SULF2 and in SULF2-expressing Huh7 cells transfected with short hairpin RNA targeting SULF2. The interaction between GPC3, SULF2, and Wnt3a was assessed by coimmunoprecipitation and flow cytometry. β-catenin-dependent transcriptional activity was assessed with the TOPFLASH (T cell factor reporter plasmid) luciferase assay. In HCC cells, SULF2 increased cell surface GPC3 and Wnt3a expression, stabilized β-catenin, and activated T cell factor transcription factor activity and expression of the Wnt/β-catenin target gene cyclin D1. Opposite effects were observed in SULF2-knockdown models. In vivo, nude mouse xenografts established from SULF2-transfected Hep3B cells showed enhanced GPC3, Wnt3a, and β-catenin levels. CONCLUSION: Together, these findings identify a novel mechanism mediating the oncogenic function of SULF2 in HCC that includes GPC3-mediated activation of Wnt signaling via the Wnt3a/glycogen synthase kinase 3 beta axis.