title: Hexokinase II in CD133+ and CD133- hepatoma BEL-7402 Cells.
abstract: 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.
authors: Department of Hepatobiliary Surgery, Peking University People's Hospital, No. 11 South Xizhimen Street, Xicheng District, Beijing, 100044, People's Republic of China.
authors: Lei
authors: L
authors: Gong
authors: Zhuqingqing
authors: Z
authors: Cui
authors: Xin
authors: X
authors: Yu
authors: Yuhua
authors: Y
authors: Wei
authors: Jirun
authors: J
authors: Peng
authors: Xisheng
authors: X
authors: Leng
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism*; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism*; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D012333:RNA, Messenger; D006593:Hexokinase