Annals of Surgical Oncology

, Volume 18, Issue 12, pp 3514–3527 | Cite as

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

  • Po-Cheng Lin
  • Shinn-Zong Lin
  • Yi-Lin Chen
  • Jeng-Shou Chang
  • Li-Ing Ho
  • Po-Yen Liu
  • Li-Fu Chang
  • Yeu-Chern Harn
  • Shee-Ping Chen
  • Li-Yi Sun
  • Pi-Chun Huang
  • Jung-Ting Chein
  • Chang-Hai Tsai
  • Chii-Wen Chou
  • Horng-Jyh Harn
  • Tzyy-Wen Chiou
Translational Research and Biomarkers

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.

Keywords

hTERT Expression hTERT mRNA Expression Brain Cancer Cell Butylidenephthalide hTERT Core Promoter 

Notes

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).

Supplementary material

10434_2011_1644_MOESM1_ESM.eps (1.4 mb)
Electronic Supplementary Material: Supplementary Figures 1 and 2 are available for this article at doi: 10.1245/s10434-011-1644-0 and are accessible for authorized users. (EPS 1430 kb)
10434_2011_1644_MOESM2_ESM.eps (697 kb)
Supplementary material 2 (EPS 697 kb)

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Copyright information

© Society of Surgical Oncology 2011

Authors and Affiliations

  • Po-Cheng Lin
    • 1
  • Shinn-Zong Lin
    • 2
    • 3
    • 4
  • Yi-Lin Chen
    • 5
  • Jeng-Shou Chang
    • 6
  • Li-Ing Ho
    • 7
  • Po-Yen Liu
    • 8
  • Li-Fu Chang
    • 1
  • Yeu-Chern Harn
    • 9
    • 10
  • Shee-Ping Chen
    • 11
  • Li-Yi Sun
    • 12
  • Pi-Chun Huang
    • 13
  • Jung-Ting Chein
    • 14
  • Chang-Hai Tsai
    • 15
    • 16
  • Chii-Wen Chou
    • 2
  • Horng-Jyh Harn
    • 17
    • 18
  • Tzyy-Wen Chiou
    • 1
  1. 1.Department of Life Science and Graduate Institute of BiotechnologyNational Dong Hwa UniversityHualienTaiwan, Republic of China
  2. 2.Center for Neuropsychiatry and Department of NeurosurgeryChina Medical University HospitalTaichungTaiwan, Republic of China
  3. 3.Graduate Institute of ImmunologyChina Medical UniversityTaichungTaiwan, Republic of China
  4. 4.Department of NeurosurgeryChina Medical University Beigan HospitalYunlinTaiwan, Republic of China
  5. 5.Graduate Institute of BiotechnologyNational Ilan UniversityIlanTaiwan, Republic of China
  6. 6.Graduate Institute of Basic Medical ScienceChina Medical UniversityTaichungTaiwan, Republic of China
  7. 7.Department of Respiratory CareVeterans General Hospital-TaipeiTaipeiTaiwan, Republic of China
  8. 8.Graduate Institute of Chinese MedicineChina Medical UniversityTaichungTaiwan, Republic of China
  9. 9.Graduate Institute of Networking and MultimediaNational Taiwain UniversityTaipeiTaiwan, Republic of China
  10. 10.The Metabolomics GroupNational Taiwan UniversityTaipeiTaiwan, Republic of China
  11. 11.Buddhist Tzu Chi Stem Cells CenterBuddhist Tzu Chi Medical CenterHualienTaiwan, Republic of China
  12. 12.Department of Biological Science and TechnologyNational Chiao Tung UniversityHsinchuTaiwan, Republic of China
  13. 13.Department of Stem Cell Applied TechnologyGwo Xi Stem Cell Applied TechnologyHsinchuTaiwan, Republic of China
  14. 14.Departmeant of Medical TechnologyChina Medical UniversityTaichungTaiwan, Republic of China
  15. 15.Department of PediatricsChina Medical University HospitalTaichungTaiwan, Republic of China
  16. 16.Department of Healthcare AdministrationAsia UniversityTaichungTaiwan, Republic of China
  17. 17.Department of PathologyChina Medical University HospitalTaichungTaiwan, Republic of China
  18. 18.Department of MedicineChina Medical UniversityTaichungTaiwan, Republic of China

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