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Ras-inducible immortalized fibroblasts: focus formation without cell cycle deregulation

Oncogene volume 21, pages 3058–3067 (2002)Cite this article

Abstract

The Ras oncogene transforms cultured murine fibroblasts into malignant, focus-forming cells, whose lack of contact inhibition is evidenced by high saturation densities. In order to investigate the reversibility of Ras transformation, as well as the kinetics of Ras-induced changes, cell lines that conditionally express oncogenic Ras were constructed. Both focus formation and increased saturation density were inducible and fully reversible. In exponentially growing cells, oncogenic Ras-expression had no effect on proliferation rates, Erk phosphorylation, or the level of cyclin D1, and Ras-induction did not confer serum-independent growth. As expected, growth to high density in uninduced cells led to quiescence with a low level of cyclin D1 and no active Erk; in this setting, Ras induction prevented full downregulation of cyclin D1 and inactivation of Erk. Our results show that Ras expression to a level sufficient for transformation leads to relatively subtle effects on known downstream targets, and that the focus formation and increased saturation density growth induced by Ras is not a result of growth factor independence.

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Acknowledgements

The excellent technical assistance of Marianne Knudsen and Rikke Ingvorsen is gratefully acknowledged. We thank Douglas Lowy for critical reading of an early version of the manuscript. This work was funded by grants 97 100 13 from the Danish Cancer Society and 9600821 from the Danish Medical Research council to BM Willumsen.

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  1. Department of Molecular Cell Biology, University of Copenhagen, DK-1353 Copenhagen, Denmark

    Kivin Jacobsen, Anja Groth & Berthe M Willumsen

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  1. Kivin Jacobsen
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  2. Anja Groth
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  3. Berthe M Willumsen
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Correspondence to Berthe M Willumsen.

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Jacobsen, K., Groth, A. & Willumsen, B. Ras-inducible immortalized fibroblasts: focus formation without cell cycle deregulation. Oncogene 21, 3058–3067 (2002). https://doi.org/10.1038/sj.onc.1205423

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