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. 1996 Apr 1;15(7):1734–1741.

Experimental elongation of telomeres extends the lifespan of immortal x normal cell hybrids.

W E Wright 1, D Brasiskyte 1, M A Piatyszek 1, J W Shay 1
PMCID: PMC450086  PMID: 8612598

Abstract

Hybrids between immortal cells that express telomerase and normal cells that lack telomerase have a limited lifespan. We demonstrate that telomerase is repressed in such hybrids. Treatment of immortal human cell lines with certain oligonucleotides resulted in telomere elongation. We took advantage of this observation to test the hypothesis that elongation of telomeres would extend the lifespan of cells in culture. An immortal human cell line was treated with an oligonucleotide to lengthen its telomeres and then was fused with mortal cells. The lifespan of these hybrid cells was longer than that of the hybrids in which telomeres had not been elongated. These observations provide the first direct evidence supporting the hypothesis that telomere length determines proliferative capacity of human cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allsopp R. C., Vaziri H., Patterson C., Goldstein S., Younglai E. V., Futcher A. B., Greider C. W., Harley C. B. Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10114–10118. doi: 10.1073/pnas.89.21.10114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry I. J., Burns J. E., Parkinson E. K. Assignment of two human epidermal squamous cell carcinomas cell lines to more than one complementation group for the immortal phenotype. Mol Carcinog. 1994 Mar;9(3):134–142. doi: 10.1002/mc.2940090305. [DOI] [PubMed] [Google Scholar]
  3. Blackburn E. H. Telomerases. Annu Rev Biochem. 1992;61:113–129. doi: 10.1146/annurev.bi.61.070192.000553. [DOI] [PubMed] [Google Scholar]
  4. Blasco M. A., Funk W., Villeponteau B., Greider C. W. Functional characterization and developmental regulation of mouse telomerase RNA. Science. 1995 Sep 1;269(5228):1267–1270. doi: 10.1126/science.7544492. [DOI] [PubMed] [Google Scholar]
  5. Broccoli D., Young J. W., de Lange T. Telomerase activity in normal and malignant hematopoietic cells. Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9082–9086. doi: 10.1073/pnas.92.20.9082. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bryan T. M., Englezou A., Gupta J., Bacchetti S., Reddel R. R. Telomere elongation in immortal human cells without detectable telomerase activity. EMBO J. 1995 Sep 1;14(17):4240–4248. doi: 10.1002/j.1460-2075.1995.tb00098.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bunn C. L., Tarrant G. M. Limited lifespan in somatic cell hybrids and cybrids. Exp Cell Res. 1980 Jun;127(2):385–396. doi: 10.1016/0014-4827(80)90443-7. [DOI] [PubMed] [Google Scholar]
  8. Chadeneau C., Hay K., Hirte H. W., Gallinger S., Bacchetti S. Telomerase activity associated with acquisition of malignancy in human colorectal cancer. Cancer Res. 1995 Jun 15;55(12):2533–2536. [PubMed] [Google Scholar]
  9. Counter C. M., Avilion A. A., LeFeuvre C. E., Stewart N. G., Greider C. W., Harley C. B., Bacchetti S. Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J. 1992 May;11(5):1921–1929. doi: 10.1002/j.1460-2075.1992.tb05245.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Counter C. M., Hirte H. W., Bacchetti S., Harley C. B. Telomerase activity in human ovarian carcinoma. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):2900–2904. doi: 10.1073/pnas.91.8.2900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feng J., Funk W. D., Wang S. S., Weinrich S. L., Avilion A. A., Chiu C. P., Adams R. R., Chang E., Allsopp R. C., Yu J. The RNA component of human telomerase. Science. 1995 Sep 1;269(5228):1236–1241. doi: 10.1126/science.7544491. [DOI] [PubMed] [Google Scholar]
  12. Gottschling D. E., Aparicio O. M., Billington B. L., Zakian V. A. Position effect at S. cerevisiae telomeres: reversible repression of Pol II transcription. Cell. 1990 Nov 16;63(4):751–762. doi: 10.1016/0092-8674(90)90141-z. [DOI] [PubMed] [Google Scholar]
  13. Greider C. W., Blackburn E. H. Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell. 1985 Dec;43(2 Pt 1):405–413. doi: 10.1016/0092-8674(85)90170-9. [DOI] [PubMed] [Google Scholar]
  14. Greider C. W. Telomeres, telomerase and senescence. Bioessays. 1990 Aug;12(8):363–369. doi: 10.1002/bies.950120803. [DOI] [PubMed] [Google Scholar]
  15. Hara E., Tsurui H., Shinozaki A., Nakada S., Oda K. Cooperative effect of antisense-Rb and antisense-p53 oligomers on the extension of life span in human diploid fibroblasts, TIG-1. Biochem Biophys Res Commun. 1991 Aug 30;179(1):528–534. doi: 10.1016/0006-291x(91)91403-y. [DOI] [PubMed] [Google Scholar]
  16. Harley C. B., Futcher A. B., Greider C. W. Telomeres shorten during ageing of human fibroblasts. Nature. 1990 May 31;345(6274):458–460. doi: 10.1038/345458a0. [DOI] [PubMed] [Google Scholar]
  17. Harley C. B. Telomere loss: mitotic clock or genetic time bomb? Mutat Res. 1991 Mar-Nov;256(2-6):271–282. doi: 10.1016/0921-8734(91)90018-7. [DOI] [PubMed] [Google Scholar]
  18. Harper J. W., Adami G. R., Wei N., Keyomarsi K., Elledge S. J. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell. 1993 Nov 19;75(4):805–816. doi: 10.1016/0092-8674(93)90499-g. [DOI] [PubMed] [Google Scholar]
  19. Harrington L. A., Greider C. W. Telomerase primer specificity and chromosome healing. Nature. 1991 Oct 3;353(6343):451–454. doi: 10.1038/353451a0. [DOI] [PubMed] [Google Scholar]
  20. Hastie N. D., Dempster M., Dunlop M. G., Thompson A. M., Green D. K., Allshire R. C. Telomere reduction in human colorectal carcinoma and with ageing. Nature. 1990 Aug 30;346(6287):866–868. doi: 10.1038/346866a0. [DOI] [PubMed] [Google Scholar]
  21. Hayashi J., Werbin H., Shay J. W. Effects of normal human fibroblast mitochondrial DNA on segregation of HeLaTG Mitochondrial DNA and on tumorigenicity of HeLaTG cells. Cancer Res. 1986 Aug;46(8):4001–4006. [PubMed] [Google Scholar]
  22. Hiyama E., Hiyama K., Yokoyama T., Matsuura Y., Piatyszek M. A., Shay J. W. Correlating telomerase activity levels with human neuroblastoma outcomes. Nat Med. 1995 Mar;1(3):249–255. doi: 10.1038/nm0395-249. [DOI] [PubMed] [Google Scholar]
  23. Hiyama E., Yokoyama T., Tatsumoto N., Hiyama K., Imamura Y., Murakami Y., Kodama T., Piatyszek M. A., Shay J. W., Matsuura Y. Telomerase activity in gastric cancer. Cancer Res. 1995 Aug 1;55(15):3258–3262. [PubMed] [Google Scholar]
  24. Hiyama K., Hiyama E., Ishioka S., Yamakido M., Inai K., Gazdar A. F., Piatyszek M. A., Shay J. W. Telomerase activity in small-cell and non-small-cell lung cancers. J Natl Cancer Inst. 1995 Jun 21;87(12):895–902. doi: 10.1093/jnci/87.12.895. [DOI] [PubMed] [Google Scholar]
  25. Kim N. W., Piatyszek M. A., Prowse K. R., Harley C. B., West M. D., Ho P. L., Coviello G. M., Wright W. E., Weinrich S. L., Shay J. W. Specific association of human telomerase activity with immortal cells and cancer. Science. 1994 Dec 23;266(5193):2011–2015. doi: 10.1126/science.7605428. [DOI] [PubMed] [Google Scholar]
  26. Kyrion G., Liu K., Liu C., Lustig A. J. RAP1 and telomere structure regulate telomere position effects in Saccharomyces cerevisiae. Genes Dev. 1993 Jul;7(7A):1146–1159. doi: 10.1101/gad.7.7a.1146. [DOI] [PubMed] [Google Scholar]
  27. Levy M. Z., Allsopp R. C., Futcher A. B., Greider C. W., Harley C. B. Telomere end-replication problem and cell aging. J Mol Biol. 1992 Jun 20;225(4):951–960. doi: 10.1016/0022-2836(92)90096-3. [DOI] [PubMed] [Google Scholar]
  28. Lindsey J., McGill N. I., Lindsey L. A., Green D. K., Cooke H. J. In vivo loss of telomeric repeats with age in humans. Mutat Res. 1991 Jan;256(1):45–48. doi: 10.1016/0921-8734(91)90032-7. [DOI] [PubMed] [Google Scholar]
  29. McEachern M. J., Blackburn E. H. A conserved sequence motif within the exceptionally diverse telomeric sequences of budding yeasts. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3453–3457. doi: 10.1073/pnas.91.8.3453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Meyne J., Ratliff R. L., Moyzis R. K. Conservation of the human telomere sequence (TTAGGG)n among vertebrates. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7049–7053. doi: 10.1073/pnas.86.18.7049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Morin G. B. Recognition of a chromosome truncation site associated with alpha-thalassaemia by human telomerase. Nature. 1991 Oct 3;353(6343):454–456. doi: 10.1038/353454a0. [DOI] [PubMed] [Google Scholar]
  32. Morin G. B. The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell. 1989 Nov 3;59(3):521–529. doi: 10.1016/0092-8674(89)90035-4. [DOI] [PubMed] [Google Scholar]
  33. Muggleton-Harris A. L., DeSimone D. W. Replicative potentials of various fusion products between WI-38 and SV40 transformed WI-38 cells and their components. Somatic Cell Genet. 1980 Nov;6(6):689–698. doi: 10.1007/BF01538968. [DOI] [PubMed] [Google Scholar]
  34. Murnane J. P., Sabatier L., Marder B. A., Morgan W. F. Telomere dynamics in an immortal human cell line. EMBO J. 1994 Oct 17;13(20):4953–4962. doi: 10.1002/j.1460-2075.1994.tb06822.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nilsson P., Mehle C., Remes K., Roos G. Telomerase activity in vivo in human malignant hematopoietic cells. Oncogene. 1994 Oct;9(10):3043–3048. [PubMed] [Google Scholar]
  36. Noda A., Ning Y., Venable S. F., Pereira-Smith O. M., Smith J. R. Cloning of senescent cell-derived inhibitors of DNA synthesis using an expression screen. Exp Cell Res. 1994 Mar;211(1):90–98. doi: 10.1006/excr.1994.1063. [DOI] [PubMed] [Google Scholar]
  37. Olovnikov A. M. A theory of marginotomy. The incomplete copying of template margin in enzymic synthesis of polynucleotides and biological significance of the phenomenon. J Theor Biol. 1973 Sep 14;41(1):181–190. doi: 10.1016/0022-5193(73)90198-7. [DOI] [PubMed] [Google Scholar]
  38. Pereira-Smith O. M., Smith J. R. Evidence for the recessive nature of cellular immortality. Science. 1983 Sep 2;221(4614):964–966. doi: 10.1126/science.6879195. [DOI] [PubMed] [Google Scholar]
  39. Pereira-Smith O. M., Smith J. R. Expression of SV40 T antigen in finite life-span hybrids of normal and SV40-transformed fibroblasts. Somatic Cell Genet. 1981 Jul;7(4):411–421. doi: 10.1007/BF01542986. [DOI] [PubMed] [Google Scholar]
  40. Pluta A. F., Zakian V. A. Recombination occurs during telomere formation in yeast. Nature. 1989 Feb 2;337(6206):429–433. doi: 10.1038/337429a0. [DOI] [PubMed] [Google Scholar]
  41. Prowse K. R., Greider C. W. Developmental and tissue-specific regulation of mouse telomerase and telomere length. Proc Natl Acad Sci U S A. 1995 May 23;92(11):4818–4822. doi: 10.1073/pnas.92.11.4818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Romero D. P., Blackburn E. H. A conserved secondary structure for telomerase RNA. Cell. 1991 Oct 18;67(2):343–353. doi: 10.1016/0092-8674(91)90186-3. [DOI] [PubMed] [Google Scholar]
  43. Ryan P. A., Maher V. M., McCormick J. J. Failure of infinite life span human cells from different immortality complementation groups to yield finite life span hybrids. J Cell Physiol. 1994 Apr;159(1):151–160. doi: 10.1002/jcp.1041590119. [DOI] [PubMed] [Google Scholar]
  44. Shay J. W., Pereira-Smith O. M., Wright W. E. A role for both RB and p53 in the regulation of human cellular senescence. Exp Cell Res. 1991 Sep;196(1):33–39. doi: 10.1016/0014-4827(91)90453-2. [DOI] [PubMed] [Google Scholar]
  45. Shay J. W., Wright W. E., Brasiskyte D., Van der Haegen B. A. E6 of human papillomavirus type 16 can overcome the M1 stage of immortalization in human mammary epithelial cells but not in human fibroblasts. Oncogene. 1993 Jun;8(6):1407–1413. [PubMed] [Google Scholar]
  46. Shay J. W., Wright W. E. Telomerase activity in human cancer. Curr Opin Oncol. 1996 Jan;8(1):66–71. doi: 10.1097/00001622-199601000-00012. [DOI] [PubMed] [Google Scholar]
  47. Singer M. S., Gottschling D. E. TLC1: template RNA component of Saccharomyces cerevisiae telomerase. Science. 1994 Oct 21;266(5184):404–409. doi: 10.1126/science.7545955. [DOI] [PubMed] [Google Scholar]
  48. Tahara H., Nakanishi T., Kitamoto M., Nakashio R., Shay J. W., Tahara E., Kajiyama G., Ide T. Telomerase activity in human liver tissues: comparison between chronic liver disease and hepatocellular carcinomas. Cancer Res. 1995 Jul 1;55(13):2734–2736. [PubMed] [Google Scholar]
  49. Vaziri H., Dragowska W., Allsopp R. C., Thomas T. E., Harley C. B., Lansdorp P. M. Evidence for a mitotic clock in human hematopoietic stem cells: loss of telomeric DNA with age. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9857–9860. doi: 10.1073/pnas.91.21.9857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Vaziri H., Schächter F., Uchida I., Wei L., Zhu X., Effros R., Cohen D., Harley C. B. Loss of telomeric DNA during aging of normal and trisomy 21 human lymphocytes. Am J Hum Genet. 1993 Apr;52(4):661–667. [PMC free article] [PubMed] [Google Scholar]
  51. Wang S. S., Zakian V. A. Telomere-telomere recombination provides an express pathway for telomere acquisition. Nature. 1990 May 31;345(6274):456–458. doi: 10.1038/345456a0. [DOI] [PubMed] [Google Scholar]
  52. Wright W. E., Shay J. W., Piatyszek M. A. Modifications of a telomeric repeat amplification protocol (TRAP) result in increased reliability, linearity and sensitivity. Nucleic Acids Res. 1995 Sep 25;23(18):3794–3795. doi: 10.1093/nar/23.18.3794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wright W. E., Shay J. W. Telomere positional effects and the regulation of cellular senescence. Trends Genet. 1992 Jun;8(6):193–197. doi: 10.1016/0168-9525(92)90232-s. [DOI] [PubMed] [Google Scholar]
  54. Xiong Y., Hannon G. J., Zhang H., Casso D., Kobayashi R., Beach D. p21 is a universal inhibitor of cyclin kinases. Nature. 1993 Dec 16;366(6456):701–704. doi: 10.1038/366701a0. [DOI] [PubMed] [Google Scholar]
  55. Yu G. L., Blackburn E. H. Developmentally programmed healing of chromosomes by telomerase in Tetrahymena. Cell. 1991 Nov 15;67(4):823–832. doi: 10.1016/0092-8674(91)90077-c. [DOI] [PubMed] [Google Scholar]
  56. Yudkin J. Sugar and disease. Nature. 1972 Sep 22;239(5369):197–199. doi: 10.1038/239197a0. [DOI] [PubMed] [Google Scholar]
  57. Zahler A. M., Prescott D. M. Telomere terminal transferase activity in the hypotrichous ciliate Oxytricha nova and a model for replication of the ends of linear DNA molecules. Nucleic Acids Res. 1988 Jul 25;16(14B):6953–6972. doi: 10.1093/nar/16.14.6953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Zakian V. A. Structure and function of telomeres. Annu Rev Genet. 1989;23:579–604. doi: 10.1146/annurev.ge.23.120189.003051. [DOI] [PubMed] [Google Scholar]
  59. de Lange T. Human telomeres are attached to the nuclear matrix. EMBO J. 1992 Feb;11(2):717–724. doi: 10.1002/j.1460-2075.1992.tb05104.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. el-Deiry W. S., Tokino T., Velculescu V. E., Levy D. B., Parsons R., Trent J. M., Lin D., Mercer W. E., Kinzler K. W., Vogelstein B. WAF1, a potential mediator of p53 tumor suppression. Cell. 1993 Nov 19;75(4):817–825. doi: 10.1016/0092-8674(93)90500-p. [DOI] [PubMed] [Google Scholar]

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