A novel class of unstable 6-thioguanine-resistant cells from dog and human kidneys.
Thioguanine-resistant primary clones were grown from single cell suspensions obtained from dog and human kidneys by enzymatic digestion. In medium containing a relatively high concentration (10 micrograms/ml) of thioguanine, thioguanine-resistant primary clones arose from each source at frequencies ranging from 10(-4) to 10(-5). A reduction in total hypoxanthine uptake was found in the thioguanine-resistant primary clones which had developed in thioguanine medium, consistent with a reduction in hypoxanthine phosphoribosyltransferase activity. When these thioguanine-resistant primary clones were subsequently grown in the absence of thioguanine and assayed for the thioguanine-resistant phenotype and hypoxanthine phosphoribosyltransferase activity, it was found that most were now thioguanine-sensitive and yielded cell-free extracts with substantial amounts of hypoxanthine phosphoribosyltransferase activity. In contrast, thioguanine-resistant human clones grown continuously in the presence of thioguanine yielded cell-free extracts with little or no detectable hypoxanthine phosphoribosyltransferase activity. Southern blot analysis demonstrated no structural alterations in the hypoxanthine phosphoribosyltransferase gene in thioguanine-resistant primary human kidney clones. These results suggest that a novel mechanism(s) for thioguanine resistance and the control of hypoxanthine phosphoribosyltransferase expression may occur in dog and human kidney cells.
Turker
M S
MS
Department of Pathology, University of Washington, Seattle 98195.
Monnat
R J
RJ
Fukuchi
K
K
Johnston
P A
PA
Ogburn
C E
CE
Weller
R E
RE
Park
J F
JF
Martin
G M
GM
D000818:Animals; D002478:Cells, Cultured; D004285:Dogs; D004351:Drug Resistance; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism*; D007668:Kidney / Q000166:cytology*; D012641:Selection, Genetic; D013866:Thioguanine / Q000494:pharmacology*
D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine