In order to investigate the expression of the glycolysis-associated genes and different proliferation between normal cells and tumor cells in response to hypoxia, and to explore the role of reactive oxygen species (ROS) in mediating metabolism of hepatoma cells and in regulating expression of glycolysis-associated genes and enzyme activity, SMMC-7721 hepatoma cell line and the normal liver cell line LO2 were taken as the investigation objects. The cell suppression ratio and surviving ratio were detected respectively under conditions of simple hypoxia, or 5 mmol/L glucose complicated with hypoxia. The mRNA levels of key enzymes that are involved in regulating glyco-metabolism were detected, including the mRNA levels of pyruvate-kinase, hexokinase, succinic dehydrogenase and isocitric dehydrogenase, and the activity of lactate dehydrognase. The proliferation gene pkb and hypoxia-inducible factor-1 (HIF-1) were detected as well. The results suggest that 1. tumor cell can endure severe hypoxia condition than normal cell, 2. under conditions of hypoxia, the strengthening of the glycolytic pathway is one of the important mechanisms to ensure the rapid proliferation of tumor cells, 3. ROS through HIF-1 mediates the changes of the gene expression of the enzyme of the glycometabolism pathway, and takes part in the regulation of the hypoxia-induced signal transduction. And the use of antioxidant will decrease the ability of tumor cell to endure hypoxia.