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Alterations in the p16INK4a and p53 tumor suppressor genes of hTERT-immortalized human fibroblasts

Oncogene volume 23, pages 3116–3121 (2004)Cite this article

Abstract

Exogenous expression of the catalytic subunit of telomerase, hTERT, in a normal human foreskin fibroblast cell strain resulted in telomerase activity and an extended proliferative lifespan prior to a period of crisis. Three immortalized cell lines with stably maintained telomere lengths were established from cells that escaped crisis. Each of these cultures underwent a significant downregulation of p16INK4a expression due to gene deletion events. One cell line also acquired mutations in both alleles of the p53 tumor suppressor gene. Downregulation of p16INK4a and loss of wild-type p53 expression occurred after escape from crisis, so these mutations are most likely not required for immortalization of these cells but rather were selected for during continuous growth in vitro. These findings emphasize the need for caution in the use of hTERT-immortalized cells in studies of normal cell biology or in tissue engineering and the need to monitor for genetic instability and the accumulation of mutations in both the p16INK4a/pRb and p53 pathways.

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Acknowledgements

This work was supported by a project grant from the National Health and Medical Research Council of Australia, and the Carcinogenesis Fellowship of the Cancer Council New South Wales. We thank Christine Smyth for FACscan analysis, Alessandra Muntoni for advice on p53 sequencing, and Helen Rizos for advice on p16INK4a PCR analysis.

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Authors and Affiliations

  1. Children's Medical Research Institute, Westmead, 214 Hawkesbury Rd, Sydney, NSW 2145, Australia

    Jane R Noble, Ze-Huai Zhong, Axel A Neumann & Roger R Reddel

  2. Sydney Cancer Centre, Medical Foundation Building, K25, University of Sydney, NSW, 2006, Australia

    John R Melki & Susan J Clark

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  1. Jane R Noble
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  2. Ze-Huai Zhong
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Correspondence to Roger R Reddel.

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Noble, J., Zhong, ZH., Neumann, A. et al. Alterations in the p16INK4a and p53 tumor suppressor genes of hTERT-immortalized human fibroblasts. Oncogene 23, 3116–3121 (2004). https://doi.org/10.1038/sj.onc.1207440

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