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Telomere dysfunction and telomerase reactivation in human leukemia cell lines after telomerase inhibition by the expression of a dominant-negative hTERT mutant

Oncogene volume 21, pages 8262–8271 (2002)Cite this article

Abstract

As activation of telomerase represents a key step in the malignant transformation process, experimental models to develop anti-telomerase drugs provide a rational basis for anticancer strategies. We analysed the short and long-term efficacy of a stably expressed dominant-negative mutant (DN) of the telomerase catalytic unit (hTERT) in UT-7 and U937 human leukemia cell lines by using an IRES-e-GFP retrovirus. As expected, telomerase inactivation resulted in drastic telomere shortening, cytogenetic instability and cell growth inhibition in all e-GFP positive DN clones after 15–35 days of culture. However, despite this initial response, 50% of e-GFP positive DN clones with short telomeres escaped from crisis after 35 days of culture and recovered a proliferation rate similar to the control cells. This rescue was associated with a telomerase reactivation inducing telomere lengthening. We identified two pathways, one involving the loss of the DN transgene expression and the other the transcriptional up-regulation of endogenous hTERT with persistence of the DN transgene expression. Although this second mechanism appears to be a very rare event (one clone), these findings suggest that genomic instability induced by short telomeres after telomerase inhibition might enhance the probability of activation or selection of telomere maintenance mechanisms dependent on hTERT transcription.

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Acknowledgements

We are grateful to S Stewart and RA Weinberg and to WS Pear for providing the constructs, to S Burns for providing the BAC clone and to F Forestier for the use of the LightCycler. We thank H Raslova and P Foliot for FISH techniques and J Dando for critically reading the manuscript. This work was supported by funds from the Institut National de la Santé et de la Recherche Médicale (INSERM), grants from the Association pour la Recherche contre le Cancer (ARC no 5606 and 7364), from Institut Federatif de Recherche of Institut Gustave Roussy (IFR 2000), from the Fondation de France (no 98001281) and from Université Paris-Sud (BQR 2000 no CR112). F Delhommeau was supported by INSERM and A Thierry by the Ligue contre le Cancer.

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Authors and Affiliations

  1. INSERM U362, PR-1, Institut Gustave Roussy, 39/53 rue Camille Desmoulins, Villejuif, 94805, France

    François Delhommeau, Antoine Thierry, Evelyne Lauret, Françoise Sainteny, William Vainchenker & Annelise Bennaceur-Griscelli

  2. Service d'Hématologie et de Cytogénétique, Hôpital de Bicêtre, 78 rue du Général Leclerc, Le Kremblin Bicêtre, 94270, France

    Danièle Feneux

  3. Department of Clinical Biology, Service d'Hématologie Biologique, Institut Gustave Roussy, 39/53 rue Camille Desmoulins, Villejuif, 94805, France

    Edwige Leclercq, Marie Hélène Courtier & Annelise Bennaceur-Griscelli

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  1. François Delhommeau
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Correspondence to Annelise Bennaceur-Griscelli.

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Delhommeau, F., Thierry, A., Feneux, D. et al. Telomere dysfunction and telomerase reactivation in human leukemia cell lines after telomerase inhibition by the expression of a dominant-negative hTERT mutant. Oncogene 21, 8262–8271 (2002). https://doi.org/10.1038/sj.onc.1206054

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