Abstract
Normal human somatic cells in culture have a limited dividing potential. This is due to DNA end replication problem, whereby telomeres shorten with each subsequent cell division. When a critical telomere length is reached cells enter senescence. To overcome this problem, immortal HeLa cell line express telomerase, an enzyme that prevents telomere shortening. Although immortal, the existence of non-dividing cells that do not incorporate 3H-thymidine over 24 h of growth has been well documented in this cell line. Using DiI labeling and high-speed cell sorting, we have separated and analyzed fractions of HeLa cells that divided vigorously as well as those that cease divisions over several days in culture. We also analyzed telomerase activity in separated fractions and surprisingly, found that the fraction of cells that divided 0–1 time over 6 days in culture have several times higher endogenous telomerase activity than the fastest dividing fraction. Additionally, the non-growing fraction regains an overall high labeling index and low SA-β-Gal activity when subcultured again. This phenomenon should be considered if telomerase inhibition is to be used as an approach to cancer therapy. In this paper we also discuss possible molecular mechanisms that underlie the observed results.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bryan TM, Englezou A, Gupta J, Bacchetti S, Reddel RR (1995) Telomere elongation in immortal human cells without detectable telomerase activity. EMBO J 14:4240–4248
Bryan TM, Englezou A, Dunham M, Reddel RR (1998) Telomere length dynamics in telomerase-positive immortal human cell populations. Exp Cell Res 239:370–378
Campisi J (1992) Gene expression in quiescent and senescent fibroblasts. Ann NY Acad Sci 21:195–201
Campisi J (2000) Cancer, aging and cellular senescence. In Vivo 14:183–188
Corey RD (2002) Telomerase inhibition, oligonucleotides, and clinical trials. Oncogene 21:631–637
Counter CM, Avilion AA, LeFeuvre CE, Stewart NG, Greider CW, Harley CB, Bacchetti S (1992) Telomere shortening associated with chromosome instability is arrested in immortal cell which express telomerase activity. EMBO J 11:1921–1929
Cristofalo VJ, Pignolo RJ, Rotenberg MO (1992) Molecular changes with in vitro cellular senescence. Ann NY Acad Sci 663:187–194
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith OM (1995) A biomarker that identifies senescent human cells in culture and aging skin in vivo. Proc Natl Acad Sci USA 92:9363–9367
Feng J, Funk WD, Wang SS, Weinrich SL, Avilion AA, Chiu CP, Adams RR, Chang E, Allsopp RC, Yu J, Le S, West MD, Harley CB, Andrews WH, Greider CW, Villeponteau B (1995) The RNA component of human telomerase. Science 269:1236–1241
Ferenac M, Polancec D, Huzak M, Pereira-Smith OM, Rubelj I (2005) Early-senescing human skin fibroblasts do not demonstrate accelerated telomere shortening. J Gerontol A Biol Sci Med Sci 60:820–829
Goodwin EC, DiMaio D (2001) Induced senescence in HeLa cervical carcinoma cells containing elevated telomerase activity and extended telomeres. Cell Growth Differ 11:525–534
Gorbunova V, Seluanov A, Pereira-Smith OM (2003) Evidence that high telomerase activity may induce a senescent-like growth arrest in human fibroblasts. J Biol Chem 287:7692–7698
Harley CB, Futcher AB, Greider CW (1990) Telomeres shorten during ageing of human fibroblast. Nature 345:458–460
Hayflick L (1965) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 37:614–636
Hayflick L, Moorhead PS (1961) The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 25:585–621
Hemann MT, Strong MA, Hao LY, Greider CW (2001) The shortest telomere, not average telomere length is critical for cell viability and chromosome stability. Cell 107:67–77
Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL, Shay JW (1994) Specific association of human telomerase activity in immortal cells and cancer. Science 266:2011–2015
Ledley FD, Soriano HE, O’Malley BW Jr, Lewis D, Darlington GJ, Finegold M (1992) DiI as a marker for cellular transplantation into solid organs. Biotechniques 13:580–587
Makarov VL, Hirose Y, Langmore JP (1997) Long G tails at both ends of human chromosomes suggest a C strand degradation mechanism for telomere shortening. Cell 88:657–666
Martinez AO, Norwood TH, Prothero JW, Martin GM (1978) Evidence for clonal attenuation of growth potential in HeLa cells. In Vitro 14:996–1002
Matsumura T, Pfendt EA, Hayflick L (1979) DNA synthesis in the human diploid cell strain WI-38 during in vitro aging: an autoradiography study. J Gerontol 34:323–327
Morin GB (1989) The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell 59:521–529
Neumann AA, Reddel RR (2002) Telomere maintenance and cancer—look, no telomerase. Nat Rev Cancer 2:879–884
Olovnikov AM (1973) The incomplete copying of template margin in enzymatic synthesis of polynucleotides and biological significance of the phenomenon. J Theor Biol 41:181–190
Pereira-Smith OM, Smith JR (1981) Expression of SV40 T antigen in finite life-span hybrids of normal and SV40-transformed fibroblasts. Somatic Cell Genet 7:411–421
Roninson IB (2003) Tumor cell senescence in cancer treatment. Can Res 63:2705–2715
Rubelj I, Venable SF, Lednicky J, Butel JS, Bilyeu T, Darlington G, Surmacz E, Campisi J, Pereira-Smith OM (1997) Loss of T-antigen sequences allows SV40-transformed human cells in crisis to acquire a senescent-like phenotype. J Gerontol A Biol Sci Med Sci 52:229–234
Rubelj I, Huzak M, Brdar B, Pereira-Smith OM (2002) A single stage mechanism controls replicative senescence through Sudden Senescence Syndrome. Biogerontology 3:213–222
Saretzki G, Von Zglinicki T (2002) Replicative aging, telomeres and oxidative stress. Ann NY Acad Sci 959:24–29
Shay JW, Bacchetti S (1997) A survey of telomerase activity in human cancer. Eur J Cancer 33:787–791
Te Poele RH, Ohorkov AL, Jardine L, Cummings J, Joel SP (2002) DNA damage is able to induce senescence in tumor cells in vitro and in vivo. Cancer Res 62:1876–1883
Touissaint O, Medrano EE, Von Zglinicki T (2000) Cellular and molecular mechanisms of stress-induced premature senescence (SIPS) of human diploid fibroblasts and melanocytes. Exp Gerontol 35:927–945
Von Zglinicki T, Nilsson E, Docke WD, Brunk UT (1995) Lipofuscin accumulation and ageing of fibroblasts. Gerontology 41:95–108
Wright WE, Piatyszek MA, Rainey WE, Byrd W, Shay JW (1996) Telomerase activity in human germline and embryonic tissues and cells. Dev Genet 18:173–179
Acknowledgments
We thank Dr. Olivia Pereira-Smith, Sam and Ann Barshop Center for Longevity and Aging Studies, University of Texas Health Science Center, San Antonio for reviewing the manuscript. This work was supported by Croatian Ministry of Science, Education and Sports grant 0098077.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ivanković, M., Ćukušić, A., Gotić, I. et al. Telomerase activity in HeLa cervical carcinoma cell line proliferation. Biogerontology 8, 163–172 (2007). https://doi.org/10.1007/s10522-006-9043-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10522-006-9043-9