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Creation of human tumour cells with defined genetic elements

Abstract

During malignant transformation, cancer cells acquire genetic mutations that override the normal mechanisms controlling cellular proliferation. Primary rodent cells are efficiently converted into tumorigenic cells by the coexpression of cooperating oncogenes1,2. However, similar experiments with human cells have consistently failed to yield tumorigenic transformants3,4,5, indicating a fundamental difference in the biology of human and rodent cells. The few reported successes in the creation of human tumour cells have depended on the use of chemical or physical agents to achieve immortalization6, the selection of rare, spontaneously arising immortalized cells7,8,9,10, or the use of an entire viral genome11. We show here that the ectopic expression of the telomerase catalytic subunit (hTERT)12 in combination with two oncogenes (the simian virus 40 large-T oncoprotein and an oncogenic allele of H-ras) results in direct tumorigenic conversion of normal human epithelial and fibroblast cells. These results demonstrate that disruption of the intracellular pathways regulated by large-T, oncogenic ras and telomerase suffices to create a human tumor cell.

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Figure 1: Expression of large-T, ras and telomerase.
Figure 2: Expression of hTERT stabilizes telomere length.
Figure 3: Expression of hTERT immortalizes large-T-expressing HEK and BJ cells.
Figure 4: Growth properties and clonality of tumorigenic cells.

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Acknowledgements

We thank M. Fleming for interpretation of tumour histology, J. Smith for the gift of early passage BJ fibroblasts, and S. Dessain, B. Elenbaas, D. Fruman, P. Steiner, S. Stewart and the members of Weinberg laboratory for helpful discussions and review of the manuscript. This work was supported in part by Merck and Co. (R.A.W.), the US NCI (R.A.W., A.S.L.), a Damon Runyon–Walter Winchell Cancer Research Foundation Postdoctoral Fellowship (W.C.H.), and a Human Frontiers Postdoctoral Fellowship (R.L.B.). C.M.C. is a Whitehead Scholar; W.C.H. is a Herman and Margaret Sokol postdoctoral fellow. R.A.W. is an American Cancer Society Research Professor and a Daniel K. Ludwig Cancer Research Professor.

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Correspondence to Robert A. Weinberg.

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Hahn, W., Counter, C., Lundberg, A. et al. Creation of human tumour cells with defined genetic elements. Nature 400, 464–468 (1999). https://doi.org/10.1038/22780

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