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© 1990 Oxford University Press

other

Introduction of the activated N-ras oncogene into human fibroblasts by retroviral vector induces morphological transformation and tumorigenicity

Anne R. Kinsella, Lucja Fiszer-Maliszewska 1, Erika L.D. Mitchell, Yaping Guo, Margaret Fox and David Scott

Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute Wilmslow Road, Manchester M20 9BX, UK

1Present address: Ludwik Hirszeld Institute of Immunology and Experimental Therapy, 53114 Wroclaw, Poland

The introduction of activated N-ras cDNA Into normal diploid human skin fibroblast cell cultures using the retroviral vector pZIPneo results in a spectrum of morphologies ranging from near normal to, in rare instances, dense piled-up colonies of morphologically transformed cells. However, none of the clones isolated were transformed as assessed by growth on agar or tumorigenicity in nude mice. Introduction of both c-myc and N-ras oncogene cDNAs into normal skin fibroblasts failed to produce transformation as assessed by growth on agar and tumorigenicity in nude mice, although c-myc infection alone conferred immortality and the resultant doubly infected cell line was immortal. Using the same construct, activated N-ras cDNA was shown to transform immortalized human fibroblasts to tuinorigenicity. However, immortalization per se was shown not to guarantee ‘co operation’ with an activated N-ras gene to give malignant transformation. Although numerical and structural chromosome aberrations (clonal and non-clonal) were observed in some of the cell strains isolated after retroviral infection, these were not directly associated with viral infecton, the presence of the oncogenes or with the morphologically transformed phenotype.


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