AIM:To evaluate the qualitative and quantitative changes in N-linked glycosylation,which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma(HCC) in rodents.METHODS:Liver tissues of(1) normal(non-tumorbearing) rats;and(2) tumor-bearing rats;were collected and were used for histological and GlycanMap~ analyses.Briefly,GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique,automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-offlight mass spectrometry and custom bioinformatics.Histopathological studies were carried out to ensure the development of HCC in the tested animals.RESULTS:The N-glycomic analysis revealed 5glycans;Glc1Man9GlcNAc2,Gal2Man3GlcNac4Fuc1Neu1,Man4GlcNac2,Gal2Man3GlcNac4Neu3OAc3,and Man3GlcNac5Fuc1,which showed significant changes in rat HCC tissues when compared with normal liver tissues.Four glycans were increased(P < 0.05) and Glc1Man9GlcNAc2 was decreased(5.89 ± 0.45 vs 3.54± 0.21,P < 0.01) in HCC tissues compared to normal liver tissues.An increase(66.5 ± 1.05 vs 62.7 ± 1.1,P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats.Importantly,HCC rats showed an increase(P < 0.05) in both tumorassociated carbohydrates and in branched glycans.The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway,which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations.CONCLUSION:The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. AIM: To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethylnitrosamine-induced hepatocellular carcinoma (HCC) in rodents. METHODS: Liver tissues of (1) normal (non-tumorbearing) rats; and 2) tumor-bearing rats; were collected and were used for histological and GlycanMap® analysis. Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96- well assay technology coupled to matrix-assisted laser desorption / ionization time-off light mass spectrometry and custom bioinformatics. Histopathological studies were carried out to ensure the development of HCC in the tested animals.RESULTS: The N-glycomic analysis revealed 5glycans; Glc1Man9GlcNAc2, Gal2Man3GlcNac4Fuc1Neu1 , Man4GlcNac2, Gal2Man3GlcNac4Neu3OAc3, and Man3GlcNac5Fuc1, which showed significant changes in rat HCC tissues when compared with normal liver tissues. Four glycans were increase (P <0.05) and Glc1Man9GlcNAc2 was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P <0.01) in HCC tissues compared to normal liver tissues. Increase (66.5 ± 1.05 vs 62.7 ± 1.1, mannose structures in HCC rats was observed compared to normal rats. Implantantly, HCC rats showed an increase (P <0.05) in both tumorassociated carbohydrates and in branched glycans. The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. CONCLUSION: The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in mannose glycans and siayl Lewis glycans common in HCC liver tissues.