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

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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.

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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

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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.

Author information

Authors and Affiliations

  1. Luoyang Orthopedic Hospital of Henan Province, Luoyang, 471000, Henan, China

    Yaojian Rao, Huijuan Liu, Chunxia Jia, Hongxing Zhang, Zesheng Cui, Ya Zhang & Jiawei Cui

  2. The orthopedic department of tongji hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology, Wuhan, China

    Wei Xiong

Authors
  1. Yaojian Rao
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  2. Wei Xiong
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  3. Huijuan Liu
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  4. Chunxia Jia
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  5. Hongxing Zhang
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  6. Zesheng Cui
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  7. Ya Zhang
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  8. Jiawei Cui
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Corresponding author

Correspondence to Yaojian Rao.

Additional information

Yaojian Rao and Wei Xiong are jointly first authors contributed equally to this work.

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Cite this article

Rao, Y., Xiong, W., Liu, H. et al. Inhibition of telomerase activity by dominant-negative hTERT retards the growth of breast cancer cells. Breast Cancer 23, 216–223 (2016). https://doi.org/10.1007/s12282-014-0553-z

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  • DOI: https://doi.org/10.1007/s12282-014-0553-z

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