Hexokinase II in CD133+ and CD133- hepatoma BEL-7402 Cells.

Hexokinase II is a key enzyme in the glycolytic pathway and CD133+ human hepatoma cells possess cancer stem cell-like properties. The expression and enzyme activity of hexokinase II in CD133+ and CD133- hepatoma cells were examined. CD133 on the surface of the hepatoma BEL-7402 cells was analyzed by flow cytometry and the cells were magnetically sorted into CD133+ and CD133- groups. CD133+ cells comprised 1.04% of the total BEL-7402 cell population. Reverse transcription-polymerase chain reaction (PCR) and quantitative real-time PCR were used to assay the expression of hexokinase II mRNA in these two groups. The level of mRNA in CD133- cells was 4.35 times greater than the level in CD133+ cells. 3,6-biphosphoglucose dehydrogenase-coupled colorimetric method and temperature-sensitive trials were applied to determine the enzyme activity of hexokinase II, which was 1.02 U/g protein in CD133+ cells and 2.47 U/g protein in CD133- cells. Hexokinase II was the major active hexokinase isoform in CD133+ cells, comprising 92.7% of the overall cellular hexokinase activity. The results indicate that hexokinase II is vitally meaningful for CD133+ hepatoma BEL-7402 cells. Hexokinase II represents a new therapeutic target for treating CD133+ hepatoma cells.