Induction of hypoxanthine phosphoribosyltransferase deficiency in human U-937 cells.
The aim of the study was to establish enzyme-deficient mutants of the human permanent cell line U-937. Following chemical mutagenesis with the use of ethyl methanesulfonate, this cell line was chronically exposed to increasing concentrations of the toxic hypoxanthine analogue 6-thioguanine. Cells surviving hypoxanthine-aminopterin-thymidine selective media were separated by glass adherence with the use of 12-O-tetradecanoyl-phorbol-13-acetate. Three mutant clones were established, which have remained hypoxanthine phosphoribosyltransferase (HPRT) deficient for a period of 7 months, as shown by indirect measurements with the use of autoradiography and scintillation counting of cells exposed to [3H]hypoxanthine. Since the phenotypic properties and growth behavior of U-937 cells have remained unaltered after the induced mutation, a highly restricted chromosomal segment coding for HPRT seems to have been mutated.
Radzun
H J
HJ
Parwaresch
M R
MR
Kreipe
H
H
D001345:Autoradiography; D002455:Cell Division; D004351:Drug Resistance; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D006801:Humans; D006822:Hybrid Cells / Q000648:ultrastructure; D019271:Hypoxanthine; D007041:Hypoxanthine Phosphoribosyltransferase / Q000172:deficiency / Q000235:genetics*; D007042:Hypoxanthines / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000648:ultrastructure; D009000:Monocytes / Q000187:drug effects* / Q000648:ultrastructure; D009154:Mutation; D010641:Phenotype; D012588:Scintillation Counting; D013866:Thioguanine / Q000494:pharmacology
D007042:Hypoxanthines; D019271:Hypoxanthine; D005020:Ethyl Methanesulfonate; D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine