Frequent activation of non-ras transforming sequences in neoplastic Syrian hamster cells initiated with chemical carcinogens.
Carcinogen-caused transformation of Syrian hamster embryo cells has been widely used as a model for experimental carcinogenesis. However, analysis of the molecular mechanisms of hamster cell transformation has been limited. To expand the understanding of the molecular basis of this system, 22 independently derived Syrian hamster neoplastic cell lines initiated with chemical carcinogens were screened for the presence of dominant transforming sequences by DNA transfection into mouse NIH3T3 cells. High molecular weight DNAs from 12 (55%) of these cell lines transformed NIH3T3 cells through serial transfection cycles. NIH3T3 transformants contained hamster-specific repetitive sequences, which co-segregated with the transformed phenotype in successive transfection rounds. Results from Southern hybridization analyses and p21ras mobility assays indicated the presence of N-ras oncogenes, presumably activated by point mutations at codon 61, in 3 of the 12 (25%) transfection positive lines, all initiated with sodium bisulfite; non-ras transforming sequences were apparently activated in the remaining 9 (75%) lines. DNA prepared from NIH3T3 transformants derived from cell line 81C39 was analysed by Southern hybridization with a battery of 38 probes including non-ras oncogenes known to score as positive in the NIH3T3 assay as well as other retroviral and mammalian oncogenes. Each probe hybridized to DNA fragments showing the mobility characteristic of NIH3T3 protooncogenes, but failed to detect homolog sequences of hamster origin, even under hybridization conditions which allowed their detection in hamster DNA. Results show that ras activation occurs at a low frequency in hamster neoplastic transformation and strongly suggest that novel transforming sequences are activated, thus validating the use of this system for investigating the role of non-ras transforming sequences in neoplasia.
Notario
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Department of Radiation Medicine, Vincent T. Lombardi Cancer Research Center, Georgetown University School of Medicine, Washington, DC 20007.
Castro
R
R
Flessate
D M
DM
Doniger
J
J
DiPaolo
J A
JA
D000818:Animals; D015139:Blotting, Southern; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D015972:Gene Expression Regulation, Neoplastic; D011905:Genes, ras; D008647:Mesocricetus; D051379:Mice; D009374:Neoplasms, Experimental / Q000139:chemically induced; D009693:Nucleic Acid Hybridization; D015689:Oncogene Protein p21(ras) / Q000032:analysis*; D013045:Species Specificity; D014162:Transfection; D014407:Tumor Cells, Cultured
D015689:Oncogene Protein p21(ras)