Isolation of a Chinese Hamster Fibroblast Mutant Defective in Hexose Transport and Aerobic Glycolysis: Its Use to Dissect the Malignant Phenotype

A procedure is described for the selection of glucose uptake mutants based upon radiation suicide of Chinese hamster fibroblasts by 2-deoxy[3H]glucose. In one of these mutants, DS 7, the ability to transport either 2-deoxyglucose or 3-O-methylglucose was decreased to one-fifth to one-fourth. Besides...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1980-05, Vol.77 (5), p.2698-2701
Main Authors: Pouysségur, Jacques, Franchi, Arlette, Salomon, Jean-Claude, Silvestre, Patrick
Format: Article
Language:English
Subjects:
Animals
Biological Transport
Cell Adhesion
Cell Division
Cell growth
Cell Line
Cell lines
Fibroblasts
Glucose - metabolism
Glycolysis
Hexoses
Mice
Mice, Nude - physiology
Mutation
Neoplasms, Experimental - metabolism
Phenotype
Phenotypes
Somatic cells
Suicide
Tumor cell line
Tumors
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Summary:A procedure is described for the selection of glucose uptake mutants based upon radiation suicide of Chinese hamster fibroblasts by 2-deoxy[3H]glucose. In one of these mutants, DS 7, the ability to transport either 2-deoxyglucose or 3-O-methylglucose was decreased to one-fifth to one-fourth. Besides this defect, DS 7 produces 1/14 th the lactic acid produced by the parent when grown on 5 mM glucose. This block in aerobic glycolysis is due to a mutation that affects the expression of the phosphoglucose isomerase gene because no isomerase activity is detected in cell extracts of DS 7. This glycolytic block makes that cell line dependent exclusively on respiration for its energy requirement. Consequently, DS 7 survives well after removal of glucose but dies quickly in the presence of oligomycin. The parental line O 23 (subclone of CCl 39) grows at low serum concentration, is anchorage-independent, and is tumorigenic in nude mice. The derived glycolytic mutant DS 7 has retained both the in vitro transformed phenotype (low serum dependence and loss of anchorage dependence) and the tumor-forming capability. The tumor cells derived from the injection of DS 7 cells have kept the original glycolytic defect. This finding suggests that the transformed properties (high hexose transport and aerobic glycolysis) that can be uncoupled from abnormal growth control are not necessary for the expression of the malignant phenotype in fibroblasts.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.77.5.2698