Journal Mobile Options
Table of Contents
Vol. 104, No. 1-4, 2004
Issue release date: 2004

Human fibroblasts expressing hTERT show remarkable karyotype stability even after exposure to ionizing radiation

Pirzio L.M. · Freulet-Marrière M.-A. · Bai Y. · Fouladi B. · Murnane J.P. · Sabatier L. · Desmaze C.
To view the fulltext, log in and/or choose pay-per-view option

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Ectopic expression of telomerase results in an immortal phenotype in various types of normal cells, including primary human fibroblasts. In addition to its role in telomere lengthening, telomerase has now been found to have various functions, including the control of DNA repair, chromatin modification, and the control of expression of genes involved in cell cycle regulation. The investigations on the long-term effects of telomerase expression in normal human fibroblast highlighted that these cells show low frequencies of chromosomal aberrations. In this paper, we describe the karyotypic stability of human fibroblasts immortalized by expression of hTERT. The ectopic overexpression of telomerase is associated with unusual spontaneous as well as radiation-induced chromosome stability. In addition, we found that irradiation did not enhance plasmid integration in cells expressing hTERT, as has been reported for other cell types. Long-term studies illustrated that human fibroblasts immortalized by telomerase show an unusual stability for chromosomes and for plasmid integration sites, both with and without exposure to ionizing radiation. These results confirm a role for telomerase in genome stabilisation by a telomere-independent mechanism and point to the possibility for utilizing hTERT-immortalized normal human cells for the study of gene targeting.   

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Blackburn EH: Switching and signaling at the telomere. Cell 106:661–673 (2001).
  2. Bortoletto E, Mognato M, Ferraro P, Canova S, Cherubini R, Celotti L, Russo A: Chromosome instability induced in the cell progeny of human T lymphocytes irradiated in G(0) with gamma-rays. Mutagenesis 16:529–537 (2001).
  3. Brachman DG, Graves D, Vokes E, Beckett M, Haraf D, Montag A, Dunphy E, Mick R, Yandell D, Weichselbaum RR: Occurrence of p53 gene deletions and human papilloma virus infection in human head and neck cancer. Cancer Res 52:4832–4836 (1992).
  4. Brenneman M, Gimble FS, Wilson JH: Stimulation of intrachromosomal homologous recombination in human cells by electroporation with site-specific endonucleases. Proc natl Acad Sci USA 93:3608–3612 (1996).
  5. Chadeneau C, Hay K, Hirte HW, Gallinger S, Bacchetti S: Telomerase activity associated with acquisition of malignancy in human colorectal cancer. Cancer Res 55:2533–2536 (1995).
  6. Chang S, Khoo CM, Naylor ML, Maser RS, DePinho RA: Telomere-based crisis: functional differences between telomerase activation and ALT in tumor progression. Genes Dev 17:88–100 (2003).
  7. Collins K, Mitchell JR: Telomerase in the human organism. Oncogene 21:564–579 (2002).
  8. Counter CM, Avilion AA, LeFeuvre CE, Stewart NG, Greider CW, Harley CB, Bacchetti S: Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11:1921–1929 (1992).
  9. Cowan JM, Beckett MA, Weichselbaum RR: Chromosome changes characterizing in vitro response to radiation in human squamous cell carcinoma lines. Cancer Res 53:5542–5547 (1993).
  10. Cui W, Aslam S, Fletcher J, Wylie D, Clinton M, Clark AJ: Stabilization of telomere length and karyotypic stability are directly correlated with the level of hTERT gene expression in primary fibroblasts. J biol Chem 277:38531–38539 (2002).
  11. de Lange T: Protection of mammalian telomeres. Oncogene 21:532–540 (2002).
  12. DePinho RA: The age of cancer. Nature 408:248–254 (2000).
  13. Desmaze C, Aurias A: In situ hybridization of fluorescent probes on chromosomes, nuclei or stretched DNA: applications in physical mapping and characterization of genomic rearrangements. Cell Mol Biol 41:925–931 (1995).
  14. Difilippantonio MJ, Petersen S, Chen HT, Johnson R, Jasin M, Kanaar R, Ried T, Nussenzweig A: Evidence for replicative repair of DNA double-strand breaks leading to oncogenic translocation and gene amplification. J Exp Med 196:469–480 (2002).
  15. Donoho G, Jasin M, Berg P: Analysis of gene targeting and intrachromosomal homologous recombination stimulated by genomic double-strand breaks in mouse embryonic stem cells. Mol Cell Biol 18:4070–4078 (1998).
  16. Ducray C, Pommier JP, Martins L, Boussin FD, Sabatier L: Telomere dynamics, end-to-end fusions and telomerase activation during the human fibroblast immortalization process. Oncogene 18:4211–4223 (1999).

    External Resources

  17. Fouladi B, Sabatier L, Miller D, Pottier G, Murnane JP: The relationship between spontaneous telomere loss and chromosome instability in a human tumor cell line. Neoplasia 2:540–554 (2000).
  18. Gisselsson D: Tumour morphology interplay between chromosome aberrations and founder cell differentiation. Histol Histopathol 17:1207–1212 (2002).
  19. Gorbunova V, Seluanov A, Pereira-Smith OM: Expression of human telomerase (hTERT) does not prevent stress-induced senescence in normal human fibroblasts but protects the cells from stress-induced apoptosis and necrosis. J biol Chem 277:38540–38549 (2002).
  20. Greider CW, Blackburn EH: Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell 43:405–413 (1985).
  21. Kadhim MA, Macdonald DA, Goodhead DT, Lorimore SA, Marsden SJ, Wright EG: Transmission of chromosomal instability after plutonium alpha-particle irradiation. Nature 355:738–740 (1992).
  22. Lengauer C, Kinzler KW, Vogelstein B: Genetic instabilities in human cancers. Nature 396:643–649 (1998).
  23. Little JB, Gorgojo L, Vetrovs H: Delayed appearance of lethal and specific gene mutations in irradiated mammalian cells. Int J Radiat Oncol Biol Phys 19:1425–1429 (1990).
  24. Lo AW, Sabatier L, Fouladi B, Pottier G, Ricoul M, Murnane JP: DNA amplification by breakage/fusion/bridge cycles initiated by spontaneous telomere loss in a human cancer cell line. Neoplasia 4:531–538 (2002).
  25. Masutomi K, Yu EY, Khurts S, Ben-Porath I, Currier JL, Metz GB, Brooks MW, Kaneko S, Murakami S, DeCaprio JA, Weinberg RA, Stewart SA, Hahn WC: Telomerase maintains telomere structure in normal human cells. Cell 114:241–253 (2003).
  26. Matzner I, Savelyeva L, Schwab M: Preferential integration of a transfected marker gene into spontaneously expressed fragile sites of a breast cancer cell line. Cancer Lett 189:207–219 (2003).
  27. Melek M, Shippen DE: Chromosome healing: spontaneous and programmed de novo telomere formation by telomerase. Bioessays 18:301–308 (1996).
  28. Miller DG, Petek LM, Russell DW: Human gene targeting by adeno-associated virus vectors is enhanced by DNA double-strand breaks. Mol Cell Biol 23:3550–3557 (2003).
  29. Mondello C, Chiesa M, Rebuzzini P, Zongaro S, Verri A, Colombo T, Giulotto E, D’Incalci M, Franceschi C, Nuzzo F: Karyotype instability and anchorage-independent growth in telomerase-immortalized fibroblasts from two centenarian individuals. Biochem Biophys Res Commun 308:914–921 (2003).
  30. Morales CP, Holt SE, Ouellette M, Kaur KJ, Yan Y, Wilson KS, White MA, Wright WE, Shay JW: Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nature Genet 21:115–118 (1999).
  31. Nakayama C, Adachi N, Koyama H: Bleomycin enhances random integration of transfected DNA into a human genome. Mutat Res 409:1–10 (1998).
  32. Natarajan AT, Darroudi F, Mullenders LH, Meijers M: The nature and repair of DNA lesions that lead to chromosomal aberrations induced by ionizing radiations. Mutat Res 160:231–236 (1986).
  33. Obe G, Pfeiffer P, Savage JR, Johannes C, Goedecke W, Jeppesen P, Natarajan AT, Martinez-Lopez W, Folle GA, Drets ME: Chromosomal aberrations: formation, identification and distribution. Mutat Res 504:17–36 (2002).
  34. Perez CF, Botchan MR, Tobias CA: DNA-mediated gene transfer efficiency is enhanced by ionizing and ultraviolet irradiation of rodent cells in vitro. I. Kinetics of enhancement. Radiat Res 104:200–213 (1985).
  35. Pfeiffer P, Goedecke W, Obe G: Mechanisms of DNA double-strand break repair and their potential to induce chromosomal aberrations. Mutagenesis 15:289–302 (2000).
  36. Porteus MH, Cathomen T, Weitzman MD, Baltimore D: Efficient gene targeting mediated by adeno-associated virus and DNA double-strand breaks. Mol Cell Biol 23:3558–3565 (2003).
  37. Rassool FV, McKeithan TW, Neilly ME, van Melle E, Espinosa R, 3rd, Le Beau MM: Preferential integration of marker DNA into the chromosomal fragile site at 3p14: an approach to cloning fragile sites. Proc natl Acad Sci USA 88:6657–6661 (1991).
  38. Rubio MA, Kim SH, Campisi J: Reversible manipulation of telomerase expression and telomere length. Implications for the ionizing radiation response and replicative senescence of human cells. J biol Chem 277:28609–28617 (2002).
  39. Rudolph KL, Millard M, Bosenberg MW, DePinho RA: Telomere dysfunction and evolution of intestinal carcinoma in mice and humans. Nature Genet 28:155–159 (2001).
  40. Sabatier L, Dutrillaux B, Martin MB: Chromosomal instability. Nature 357:548 (1992).
  41. Schwartz JL, Jordan R, Evans HH, Lenarczyk M, Liber H: The TP53 dependence of radiation-induced chromosome instability in human lymphoblastoid cells. Radiat Res 159:730–736 (2003).
  42. Sharma GG, Gupta A, Wang H, Scherthan H, Dhar S, Gandhi V, Iliakis G, Shay JW, Young CS, Pandita TK: hTERT associates with human telomeres and enhances genomic stability and DNA repair. Oncogene 22:131–146 (2003).
  43. Slijepcevic P, Bryant PE: Chromosome healing, telomere capture and mechanisms of radiation-induced chromosome breakage. Int J Radiat Biol 73:1–13 (1998).
  44. Smith LL, Coller HA, Roberts JM: Telomerase modulates expression of growth-controlling genes and enhances cell proliferation. Nature Cell Biol 5:474–479 (2003).
  45. Sprung CN, Afshar G, Chavez EA, Lansdorp P, Sabatier L, Murnane JP: Telomere instability in a human cancer cell line. Mutat Res 429:209–223. (1999a).
  46. Sprung CN, Reynolds GE, Jasin M, Murnane JP: Chromosome healing in mouse embryonic stem cells. Proc natl Acad Sci USA 96:6781–6786. (1999b).
  47. Sprung CN, Sabatier L, Murnane JP: Telomere dynamics in a human cancer cell line. Exp Cell Res 247:29–37 (1999c).
  48. Stevens CW, Zeng M, Cerniglia GJ: Ionizing radiation greatly improves gene transfer efficiency in mammalian cells. Hum Gene Ther 7:1727–1734 (1996).
  49. Stevens CW, Cerniglia GJ, Giandomenico AR, Koch CJ: DNA damaging agents improve stable gene transfer efficiency in mammalian cells. Radiat Oncol Investig 6:1–9 (1998).
  50. Stevens CW, Stamato TD, Mauldin SK, Getts RC, Zeng M, Cerniglia GJ: Radiation-induced recombination is dependent on Ku80. Radiat Res 151:408–413 (1999).
  51. Stevens CW, Puppi M, Cerniglia GJ: Time-dose relationships in radiation-enhanced integration. Int J Radiat Biol 77:841–846 (2001).
  52. Stewart SA, Hahn WC, O’Connor BF, Banner EN, Lundberg AS, Modha P, Mizuno H, Brooks MW, Fleming M, Zimonjic DB, Popescu NC, Weinberg RA: Telomerase contributes to tumorigenesis by a telomere length-independent mechanism. Proc natl Acad Sci USA 99:12606–12611 (2002).
  53. Tang R, Cheng AJ, Wang JY, Wang TC: Close correlation between telomerase expression and adenomatous polyp progression in multistep colorectal carcinogenesis. Cancer Res 58:4052–4054 (1998).
  54. Zeng M, Cerniglia GJ, Eck SL, Stevens CW: High-efficiency stable gene transfer of adenovirus into mammalian cells using ionizing radiation. Hum Gene Ther 8:1025–1032 (1997).
  55. Zhu J, Wang H, Bishop JM, Blackburn EH: Telomerase extends the lifespan of virus-transformed human cells without net telomere lengthening. Proc natl Acad Sci USA 96:3723–3728 (1999).

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50