Isolation of mutants temperature-sensitive for expression of the transformed state from chemically transformed C3H/10T1/2 cells

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Abstract
58 MCA Cl 16 is an oncogenic methylcholanthrene-transformed variant of the non-transformed mouse embryo fibroblast cell line, C3H/10T1/2 Cl 8. Using two different protocols, we have isolated six temperature-sensitive mutants from N-methyl-N'-nitro-N-nitrosoguanidine treated cultures of 58 MCA CI 16. C3H/10T1/2 Cl 8, 58 MCA Cl 16 and the six mutant lines were characterized with respect to several properties associated with the transformed state: morphology, saturation density, anchorage independence, cell surface morphology and growth in medium containing 1% fetal calf serum. In general, C3H/10T1/2 cells behaved as non-transformed, whether grown at 33°C or 39.5°C. The transformed parental line and all six mutants behaved as transformed cells at 33°C. At 39.5°C, only the parental transformed line retained the transformed phenotype. Three of the mutants revert towards non-transformed behavior at 39.5°C for all of the properties tested. The remaining mutants are temperature-sensitive for some, but not all, transformed characteristics. Thus, while the expression of these transformed properties is sometimes coupled, we have been able to dissociate the expression of traits such as saturation density, anchorage independence and transformed morphology from each other. These mutants should prove to be valuable tools in the study of the mechanisms which underly the expression of the chemically-induced transformed state.