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  • Title: Activation of the c-Ha-ras-1 proto-oncogene by methylation in vitro with alpha-acetoxy-N-nitrosodimethylamine.
    Author: Hirani-Hojatti S, Milligan JR, Kovnat A, Brown K, Archer MC.
    Journal: Mol Carcinog; 1989; 2(2):101-6. PubMed ID: 2669817.
    Abstract:
    alpha-Acetoxy-N-nitrosodimethylamine, an activated derivative of the carcinogen N-nitrosodimethylamine, methylated in vitro a plasmid containing the human c-Ha-ras-1 proto-oncogene, resulting in the generation of a transforming oncogene, assayed by transfection into NIH 3T3 cells. The resulting transformed cells were tumorigenic and metastatic in immune-deprived mice. Further transfection using tumor DNA led to the formation of three secondary NIH 3T3 transformants. DNA from these secondary transformants contained human ras gene sequences. Two of the three secondary transformants contained G----A mutations at guanine 35 in codon 12, and the third secondary transformant retained the wild-type sequence at codons 12, and 61. For the latter, the activating mutation was not determined. These results demonstrate that a simple methylating agent can activate a normal human ras proto-oncogene to a transforming oncogene.
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