Breast Cancer

, Volume 23, Issue 2, pp 216–223 | Cite as

Inhibition of telomerase activity by dominant-negative hTERT retards the growth of breast cancer cells

  • Yaojian RaoEmail author
  • Wei Xiong
  • Huijuan Liu
  • Chunxia Jia
  • Hongxing Zhang
  • Zesheng Cui
  • Ya Zhang
  • Jiawei Cui
Original Article



Telomerase, a ribonucleoprotein enzyme mainly consisted of a catalytic protein subunit human telomerase reverse transcriptase (hTERT) and a human telomerase RNA component, is responsible for maintaining telomeres. Telomerase over-expression correlates significantly with tumors and is a prognostic marker. However, telomerase over-expression in breast cancers and the effect of telomerase inhibition as a candidate cancer therapy are unknown.


We used the dominant-negative mutant of hTERT (DN-hTERT) to inhibit telomerase activity on human breast adenocarcinoma cell line MCF-7 by transfection. Telomeric repeat amplification protocol assays and real-time quantitative RT-PCR were performed to investigate telomerase activity as well as expression of hTERT. Telomere length was measured by the flow-fluorescence in situ hybridization assay. Cell proliferation was assessed by the WST-8 assay, and apoptosis was evaluated by flow cytometry. The tumor formation ability of MCF-7 cells was investigated by transplanting cells subcutaneously into BALB/c nude mice.


Ectopic expression of DN-hTERT caused dramatically inhibition of telomerase activity and reduction of telomere length. Telomerase inhibition induced growth arrest and apoptosis of MCF7 cells in vitro and loss of tumorigenic properties in vivo.


This study shows that telomerase inhibition by DN-hTERT can effectively inhibit the cell viability and tumorigenicity of MCF7 cells and is an attractive approach for breast cancer therapy.


MCF-7 Telomerase activity DN-hTERT Breast cancer 



Dominant negative-human telomerase reverse transcriptase


Human telomerase RNA


Human telomerase reverse transcriptase


Polymerase chain reaction


Telomeric repeat amplification protocol



This work was financially supported by the National Science Foundation of China (No. 30500596).

Conflict of interest

The authors declare that they have no competing interests.


  1. 1.
    DeSantis C, Siegel R, Bandi P, Jemal A. Breast cancer statistics, 2011. CA Cancer J Clin. 2011;61:409–18.CrossRefPubMedGoogle Scholar
  2. 2.
    Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61:69–90.CrossRefPubMedGoogle Scholar
  3. 3.
    Coughlin SS, Ekwueme DU. Breast cancer as a global health concern. Cancer Epidemiol. 2009;33:315–8.CrossRefPubMedGoogle Scholar
  4. 4.
    Moyzis RK, Buckingham JM, Cram LS, Dani M, Deaven LL, Jones MD, et al. A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes. Proc Natl Acad Sci USA. 1988;85:6622–6.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Zakian VA. Telomeres: beginning to understand the end. Science. 1995;270:1601–7.CrossRefPubMedGoogle Scholar
  6. 6.
    Grandin N, Charbonneau M. Protection against chromosome degradation at the telomeres. Biochimie. 2008;90:41–59.CrossRefPubMedGoogle Scholar
  7. 7.
    Cohen SB, Graham ME, Lovrecz GO, Bache N, Robinson PJ, Reddel RR. Protein composition of catalytically active human telomerase from immortal cells. Science. 2007;315:1850–3.CrossRefPubMedGoogle Scholar
  8. 8.
    Shay JW, Wright WE. Senescence and immortalization: role of telomeres and telomerase. Carcinogenesis. 2005;26:867–74.CrossRefPubMedGoogle Scholar
  9. 9.
    Hiyama E, Hiyama K, Yokoyama T, Shay JW. Immunohistochemical detection of telomerase (hTERT) protein in human cancer tissues and a subset of cells in normal tissues. Neoplasia. 2001;3:17–26.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, et al. Specific association of human telomerase activity with immortal cells and cancer. Science. 1994;266:2011–5.CrossRefPubMedGoogle Scholar
  11. 11.
    Carey LA, Kim NW, Goodman S, Marks J, Henderson G, Umbricht CB, et al. Telomerase activity and prognosis in primary breast cancers. J Clin Oncol. 1999;17:3075–81.PubMedGoogle Scholar
  12. 12.
    Hahn WC, Stewart SA, Brooks MW, York SG, Eaton E, Kurachi A, et al. Inhibition of telomerase limits the growth of human cancer cells. Nat Med. 1999;5:1164–70.CrossRefPubMedGoogle Scholar
  13. 13.
    Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH, et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science. 1996;272:263–7.CrossRefPubMedGoogle Scholar
  14. 14.
    Wyatt HD, Tsang AR, Lobb DA, Beattie TL. Human telomerase reverse transcriptase (hTERT) Q169 is essential for telomerase function in vitro and in vivo. PLoS One. 2009;4:e7176.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Wang F, Zhou H, Xia X, Sun Q, Wang Y, Cheng B. Activated Notch signaling is required for hepatitis B virus X protein to promote proliferation and survival of human hepatic cells. Cancer Lett. 2010;298:64–73.CrossRefPubMedGoogle Scholar
  16. 16.
    Martens UM, Brass V, Sedlacek L, Pantic M, Exner C, Guo Y, et al. Telomere maintenance in human B lymphocytes. Br J Haematol. 2002;119:810–8.CrossRefPubMedGoogle Scholar
  17. 17.
    Guide for the care and use of laboratory animals. Guide for the care and use of laboratory animals. 8th ed. Washington: National Academies Press; 2011.Google Scholar
  18. 18.
    Chan SR, Blackburn EH. Telomeres and telomerase. Philos Trans R Soc Lond B Biol Sci. 2004;359:109–21.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Rhyu MS. Telomeres, telomerase, and immortality. J Natl Cancer Inst. 1995;87:884–94.CrossRefPubMedGoogle Scholar
  20. 20.
    Neidle S, Parkinson G. Telomere maintenance as a target for anticancer drug discovery. Nat Rev Drug Discov. 2002;1:383–93.CrossRefPubMedGoogle Scholar
  21. 21.
    Shay JW, Bacchetti S. A survey of telomerase activity in human cancer. Eur J Cancer. 1997;33:787–91.CrossRefPubMedGoogle Scholar
  22. 22.
    Shay JW, Zou Y, Hiyama E, Wright WE. Telomerase and cancer. Hum Mol Genet. 2001;10:677–85.CrossRefPubMedGoogle Scholar
  23. 23.
    Meyerson M. Role of telomerase in normal and cancer cells. J Clin Oncol. 2000;18:2626–34.PubMedGoogle Scholar
  24. 24.
    Parkinson EK, Minty F. Anticancer therapy targeting telomeres and telomerase: current status. BioDrugs. 2007;21:375–85.CrossRefPubMedGoogle Scholar
  25. 25.
    Sachsinger J, Gonzalez-Suarez E, Samper E, Heicappell R, Muller M, Blasco MA. Telomerase inhibition in RenCa, a murine tumor cell line with short telomeres, by overexpression of a dominant negative mTERT mutant, reveals fundamental differences in telomerase regulation between human and murine cells. Cancer Res. 2001;61:5580–6.PubMedGoogle Scholar
  26. 26.
    Chin K, de Solorzano CO, Knowles D, Jones A, Chou W, Rodriguez EG, et al. In situ analyses of genome instability in breast cancer. Nat Genet. 2004;36:984–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Hochreiter AE, Xiao H, Goldblatt EM, Gryaznov SM, Miller KD, Badve S, et al. Telomerase template antagonist GRN163L disrupts telomere maintenance, tumor growth, and metastasis of breast cancer. Clin Cancer Res. 2006;12:3184–92.CrossRefPubMedGoogle Scholar
  28. 28.
    Joseph I, Tressler R, Bassett E, Harley C, Buseman CM, Pattamatta P, et al. The telomerase inhibitor imetelstat depletes cancer stem cells in breast and pancreatic cancer cell lines. Cancer Res. 2010;70:9494–504.CrossRefPubMedGoogle Scholar
  29. 29.
    Price JE, Polyzos A, Zhang RD, Daniels LM. Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice. Cancer Res. 1990;50:717–21.PubMedGoogle Scholar

Copyright information

© The Japanese Breast Cancer Society 2014

Authors and Affiliations

  • Yaojian Rao
    • 1
    Email author
  • Wei Xiong
    • 2
  • Huijuan Liu
    • 1
  • Chunxia Jia
    • 1
  • Hongxing Zhang
    • 1
  • Zesheng Cui
    • 1
  • Ya Zhang
    • 1
  • Jiawei Cui
    • 1
  1. 1.Luoyang Orthopedic Hospital of Henan ProvinceLuoyangChina
  2. 2.The orthopedic department of tongji hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology Wuhan China

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