Document Detail


Kinetics of transport and phosphorylation of glucose in cancer cells.
MedLine Citation:
PMID:  19681047     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Metabolic control analysis of tumor glycolysis has indicated that hexokinase (HK) and glucose transporter (GLUT) exert the main flux control (71%). To understand why they are the main controlling steps, the GLUT and HK kinetics and the contents of GLUT1, GLUT2, GLUT3, GLUT4, HKI, and HKII were analyzed in rat hepatocarcinoma AS-30D and HeLa human cervix cancer. An improved protocol to determine the kinetic parameters of GLUT was developed with D-[2-(3)H-glucose] as physiological substrate. Kinetic analysis revealed two components at low- and high-glucose concentrations in both tumor cells. At low glucose and 37 degrees C, the V(max) was 55 +/- 20 and 17.2 +/- 6 nmol (min x mg protein)(-1), whereas the K(m) was 0.52 +/- 0.7 and 9.3 +/- 3 mM for hepatoma and HeLa cells, respectively. GLUT activity was partially inhibited by cytochalasin B (IC(50) = 0.44 +/- 0.1; K(i) = 0.3 +/- 0.1 microM) and phloretin (IC(50) = 8.7 microM) in AS-30D hepatocarcinoma. At physiological glucose, GLUT1 and GLUT3 were the predominant active isoforms in HeLa cells and AS-30D cells, respectively. HK activity in HeLa cells was much lower (60 mU/mg protein) than that in AS-30D cells (700 mU/mg protein), but both HKs were strongly inhibited by G6P. HKII was the predominant isoform in AS-30D carcinoma and HeLa cells. The much lower GLUT V(max) and catalytic efficiency (V(max)/K(m)) values in comparison to those of G6P-sensitive HK suggested the transporter exerts higher control on the glycolytic flux than HK in cancer cells. Thus, GLUT seems a more adequate therapeutic target.
Authors:
Sara Rodríguez-Enríquez; Alvaro Marín-Hernández; Juan Carlos Gallardo-Pérez; Rafael Moreno-Sánchez
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of cellular physiology     Volume:  221     ISSN:  1097-4652     ISO Abbreviation:  J. Cell. Physiol.     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-10-01     Completed Date:  2009-11-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0050222     Medline TA:  J Cell Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  552-9     Citation Subset:  IM    
Affiliation:
Departamento de Bioquímica, Instituto Nacional de Cardiología Ignacio Chávez, Tlalpan, México City, Mexico. sara.rodriguez@cardiologia.org.mx
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Transport / drug effects
Catalysis / drug effects
Cell Line, Tumor
Cell Proliferation
Cold Temperature
Cytochalasin B / pharmacology
Cytosol / drug effects,  metabolism
Female
Glucose / metabolism*
Glucose Transport Proteins, Facilitative / antagonists & inhibitors,  metabolism
Glucose-6-Phosphate / pharmacology
Glycolysis / physiology
Hela Cells
Hexokinase / analysis,  metabolism
Humans
Insulin / pharmacology
Isoenzymes / metabolism
Kinetics
Mitochondria / drug effects,  metabolism
Neoplasms / metabolism*
Phloretin / pharmacology
Phosphorylation
Rats
Rats, Wistar
Chemical
Reg. No./Substance:
0/Glucose Transport Proteins, Facilitative; 0/Isoenzymes; 11061-68-0/Insulin; 14930-96-2/Cytochalasin B; 50-99-7/Glucose; 56-73-5/Glucose-6-Phosphate; 60-82-2/Phloretin; EC 2.7.1.1/Hexokinase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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