ObjectiveTo summarize the characteristics of hexosamine biosynthesis pathway (HBP) and O-glycosylation and their roles in metabolism of hepatocellular carcinoma. MethodTo review the current literatures on the role of HBP and O-glycosylation in tumors, especially in hepatocellular carcinoma. ResultsThere was metabolic reprogramming in hepatocellular carcinoma cells, and the HBP was a branch of glycolysis pathway, which played an important role in tumorigenesis, development, and metastasis. HBP provided uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc) for O-glycosylation, UDP-GlcNAc was a substrate for OGT, participating in O-glycosylation. O-glycosylation was a type of posttranslational modification that regulates the biological behavior of tumor cells by glycosylation of target proteins in tumor cells. ConclusionHBP and O-glycosylation can be used as intervention targets in the treatment of hepatocellular carcinoma, which provides a potential method for scientific prevention and treatment of hepatocellular carcinoma.