Resistance to methyl mercaptopurine riboside in cultured hamster cells. Preliminary characterization of resistant cells and conditions affecting their selection in quantitative mutation studies.
Cells resistant to high concentrations of methyl mercaptopurine riboside (MMPR), an anlogue of adenosine, were found at a frequency of about 6 x 10(-6) per viable cell in untreated cultures of V79 Chinese hamster cells. Resistant cells were selected less efficiently if purines were added to the MMPR-medium, but high cell densities had little effect upon selection. 6 independently-isolated spontaneous MMPR-resistant sublines were characterized by their resistance to the toxicity of different purines, rate of purine excretion, incorporation of radioactive adenosine, electrophoresis of cell extracts, and expression of resistance in hybrids to an MMPR-sensitive line. 5 of these sublines showed recessive expression of MMPR-resistance in hybrids and had characteristics consistent with loss of adenosine kinase activity, while the remaining subline was much less resistant to MMPR and showed semi-dominant expression of resistance without loss of adenosine kinase activity. Cells with high resistance to MMPR were not found in a "tetraploid" (hybrid) V79 line or in freshly-isolated human cell cultures, but occurred at a comparable frequency to V79 in another commonly-used aneuploid hamster line, CHO-K1. The frequency of MMPR-resistant cells in V79 cultures was increased to a similar extent by treatment with gamma-rays or with ethyl methanesulphonate, providing a suitable post-treatment interval was allowed for the expression of resistance. A genetic interpretation of these data is given in which it is proposed that resistance most usually arises through mutation of an autosomally-linked gene of which one copy has been inactivated or lost in V79 and in CHO-K1 cells. In comparison to published data on the selection of "mutants" resistant to 6-thioguanine, it is argued that MMPR could be as useful a selective agent as thioguanine and may select a different range of types of mutagenic event.
D000248:Adenosine Kinase / Q000235:genetics; D000818:Animals; D002459:Cell Fusion; D002460:Cell Line; D002478:Cells, Cultured; D006224:Cricetinae; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004351:Drug Resistance; D005020:Ethyl Methanesulfonate; D007288:Inosine / Q000031:analogs & derivatives*; D008778:Methylthioinosine / Q000494:pharmacology*; D009153:Mutagens; D011839:Radiation, Ionizing
D009153:Mutagens; D008778:Methylthioinosine; D007288:Inosine; D005020:Ethyl Methanesulfonate; D000248:Adenosine Kinase