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Butylidenephthalide Suppresses Human Telomerase Reverse Transcriptase (TERT) in Human Glioblastomas

  • Translational Research and Biomarkers
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Abstract

Background

Telomerase is widely expressed in most human cancers, but is almost undetectable in normal somatic cells and is therefore a potential drug target. Using the human telomerase promoter platform, the naturally occurring compound butylidenephthalide (BP) was selected for subsequent investigation of antitumor activity in vitro and in vivo.

Methods

We treated human glioblastoma cells with BP and found a dose-dependent decrease in human telomerase reverse transcriptase (hTERT) mRNA expression and a concomitant increase in p16 and p21 expression. Because c-Myc and Sp1 are involved in transcriptional regulation of hTERT, the effect of BP on c-Myc and Sp1 expression was examined.

Results

Using electrophoretic mobility shift assays and western blotting, we showed that BP represses hTERT transcriptional activity via downregulation of Sp1 expression. Using the telomerase repeat amplification protocol, an association between BP concentration and suppression of telomerase activity, induction of human glioblastoma senescence, and inhibition of cellular proliferation was identified. This was supported by a mouse xenograft model, in which BP repressed telomerase and inhibited tumor proliferation, resulting in tumor senescence. Overexpression of hTERT restored telomerase activity in human glioblastoma cells and overcame replicative senescence.

Conclusions

These findings suggest that BP inhibits proliferation and induces senescence in human glioblastomas by downregulating hTERT expression and consequently telomerase activity. This is the first study to describe regulation of telomerase activity by BP in human glioblastomas.

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Acknowledgment

We thank Robert A. Weinberg for the pCI neo-hEST2-HA (Addgene plasmid 1782) construct. Grant: This work was supported by grants from the National Science Council of the Republic of China (96-2320-B-039-044-MY3), from the Center for Neuropsychiatry, China Medical University and Hospital, Taichung, Taiwan, and from China Medical University and Hospital, Taichung, Taiwan (DMR-99-076). This study is supported in part by Taiwan Department of Health Clinical Trial and Research Center of Excellence (DOH99-TD-B-111-004).

Author information

Authors and Affiliations

  1. Department of Life Science and Graduate Institute of Biotechnology, National Dong Hwa University, Hualien, Taiwan, Republic of China

    Po-Cheng Lin PhD, Li-Fu Chang MS & Tzyy-Wen Chiou PhD

  2. Center for Neuropsychiatry and Department of Neurosurgery, China Medical University Hospital, Taichung, Taiwan, Republic of China

    Shinn-Zong Lin MD, PhD & Chii-Wen Chou MD

  3. Graduate Institute of Immunology, China Medical University, Taichung, Taiwan, Republic of China

    Shinn-Zong Lin MD, PhD

  4. Department of Neurosurgery, China Medical University Beigan Hospital, Yunlin, Taiwan, Republic of China

    Shinn-Zong Lin MD, PhD

  5. Graduate Institute of Biotechnology, National Ilan University, Ilan, Taiwan, Republic of China

    Yi-Lin Chen PhD

  6. Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan, Republic of China

    Jeng-Shou Chang MS

  7. Department of Respiratory Care, Veterans General Hospital-Taipei, Taipei, Taiwan, Republic of China

    Li-Ing Ho MD

  8. Graduate Institute of Chinese Medicine, China Medical University, Taichung, Taiwan, Republic of China

    Po-Yen Liu MS

  9. Graduate Institute of Networking and Multimedia, National Taiwain University, Taipei, Taiwan, Republic of China

    Yeu-Chern Harn BS

  10. The Metabolomics Group, National Taiwan University, Taipei, Taiwan, Republic of China

    Yeu-Chern Harn BS

  11. Buddhist Tzu Chi Stem Cells Center, Buddhist Tzu Chi Medical Center, Hualien, Taiwan, Republic of China

    Shee-Ping Chen PhD

  12. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, Republic of China

    Li-Yi Sun PhD

  13. Department of Stem Cell Applied Technology, Gwo Xi Stem Cell Applied Technology, Hsinchu, Taiwan, Republic of China

    Pi-Chun Huang MS

  14. Departmeant of Medical Technology, China Medical University, Taichung, Taiwan, Republic of China

    Jung-Ting Chein BS

  15. Department of Pediatrics, China Medical University Hospital, Taichung, Taiwan, Republic of China

    Chang-Hai Tsai MD, PhD

  16. Department of Healthcare Administration, Asia University, Taichung, Taiwan, Republic of China

    Chang-Hai Tsai MD, PhD

  17. Department of Pathology, China Medical University Hospital, Taichung, Taiwan, Republic of China

    Horng-Jyh Harn MD, PhD

  18. Department of Medicine, China Medical University, Taichung, Taiwan, Republic of China

    Horng-Jyh Harn MD, PhD

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  1. Po-Cheng Lin PhD
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Corresponding author

Correspondence to Tzyy-Wen Chiou PhD.

Additional information

P-C. Lin and S-Z. Lin contributed equally to this work.

H-J. Harn and T-W. Chiou contributed equally to this work.

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Lin, PC., Lin, SZ., Chen, YL. et al. Butylidenephthalide Suppresses Human Telomerase Reverse Transcriptase (TERT) in Human Glioblastomas. Ann Surg Oncol 18, 3514–3527 (2011). https://doi.org/10.1245/s10434-011-1644-0

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