Effects of H-ras and v-sis overexpression on N-acetylglucosaminyltransferase V and metastasis-related phenotypes in human hepatocarcinoma cells.

Oncogenes and N-acetylglucosaminyltransferase (GnT-V) are both commonly associated with carcinogenesis and metastasis. In order to elucidate the relationship between oncogenes and GnT-V, two oncogenes, H-ras and v-sis/PDGF (platelet-derived growth factor), were selected, and the effects of their overexpression on GnT-V in 7721 human hepatocarcinoma cells were investigated. The results showed that the over expression of H-ras or v-sis/PDGF-B up-regulated the activities of GnT-V to various degrees in the transfected cells. In H-ras- and PDGF-B-overexpressing cells, the activity of GnT-V was up-regulated to double the normal value. The transient expression of v-sis, which produces a protein almost identical to PDGF-B, stimulated the GnT-V activity by 80.3%, and the effect was more pronounced (increased by 182.5%) in 7721 cells with stable expression of v-sis. The stimulating effect was entirely abolished by treatment with PDGF-B antibody. The staining of asparagine-linked glycans (N-glycans) in the H-ras- and v-sis-overexpressing 7721 cells was intensified when horseradish peroxidase-labeled leucoagglutinating phytohemogglutinin was used as a probe, indicating the increased content of beta1,6GlcNAc branching on the N-glycans. The enhancement of GnT-V mRNA expression was also observed in H-ras- and v-sis- overexpressing cells, indicating that H-ras and v-sis regulated GnT-V via the transcription of GnT-V mRNA and the synthesis of GnT-V protein. The cells overexpressing H-ras and v-sis displayed some changes in metastasis-related phenotypes, including acceleration of cell growth, decline of cell adhesion to fibronectin, and an increase of cell adhesion to laminin, as well as increased invasiveness through Matrigel. These results indicated that the alteration of cell adhesion and invasion induced by oncogenes is closely related to the up-regulation of GnT-V activity and its product, beta1,6GlcNAc branching in N-glycans on the cell surface.