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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

Abstract

Background

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.

Methods

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.

Results

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.

Conclusion

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.

Keywords

MCF-7 Telomerase activity DN-hTERT Breast cancer 

Abbreviations

DN-hTERT

Dominant negative-human telomerase reverse transcriptase

hTR

Human telomerase RNA

hTERT

Human telomerase reverse transcriptase

PCR

Polymerase chain reaction

TRAP

Telomeric repeat amplification protocol

Notes

Acknowledgments

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.

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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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