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  • Title: Activation of H-ras oncogenes in preneoplastic mouse mammary tissues.
    Author: Kumar R, Medina D, Sukumar S.
    Journal: Oncogene; 1990 Aug; 5(8):1271-7. PubMed ID: 2118247.
    Abstract:
    The mammary hyperplastic outgrowth (HOG) line C4, resulted from serial transplantation of a hyperplastic alveolar nodule which arose in a dimethylbenz(a)anthracene (DMBA) treated mouse. The immortalized C4 outgrowth line, on transplantation into syngeneic mice, develops as preneoplastic, hyperplastic outgrowths and subsequently into malignant carcinomas after a long latent period (greater than 6 months). Treatment of mice carrying C4 HOG transplants with DMBA resulted in a reduced latent period for tumor development (less than 3 months) and an increased tumor incidence. DNA's from C4 HOGs and mammary carcinomas of untreated as well as DMBA-treated mice were analyzed for the presence of oncogenes by the NIH3T3 focus forming assay. Transforming H-ras genes were detected in two of 6 preneoplastic HOGs and 10 of 12 carcinomas from DMBA-treated mice. DNAs from neither the HOGs nor the tumors from untreated mice were positive in this assay. The H-ras locus was then directly examined in the 61st codon by in vitro amplification of each of the tissue DNAs using PCR. The location of the activating mutation was determined by hybridization of amplified DNA to mixed sequence oligonucleotide probes. The specific nature of the mutation was defined by RFLPs using XbaI, TaqI and Sau96I restriction enzymes. Six of the H-ras oncogenes in DMBA-promoted tumors were activated by commonly observed A to T transversions at the 61st codon, while five (including an additional tumor with H-ras oncogene revealed by PCR analysis) contained novel A to G transitions. The H-ras oncogene in one DMBA-treated HOG sample was activated by A to T while the second contained an A to G mutations, representative of both modes of mutational activation involved in this model of mammary tumorigenesis. In summary, DMBA-induced point mutated H-ras oncogenes appear to potentiate the progression of hyperplastic outgrowths (HOG) to mammary carcinomas.
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