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Epidermal growth factor activates telomerase activity by direct binding of Ets-2 to hTERT promoter in lung cancer cells

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Tumor Biology

Abstract

Growth signals are directly or indirectly involved in telomerase regulation. In this study, we investigated molecular mechanisms of the effect of EGF (epidermal growth factor) on regulating hTERT (human telomerase reverse transcriptase) expression. To elucidate specific transcription factors involved in EGF-stimulated hTERT transcription in A549 and H1299 lung cancer cells, transcription factors drives hTERT promoter activity, such as Myc, Mad, and Ets-2, was evaluated on luciferase reporter assay. The upregulation of hTERT promoter by Ets-2 and Myc were abolished by Mad. Using DAPA (DNA affinity precipitation assay), Ets-2 binding to SNP (T) was stronger than Ets-2 binding to SNP (C) at −245 bp upstream of the transcription start site within the core promoter of hTERT. Ets-2 silence by siRNA decreased hTERT expression at mRNA and protein levels. The regulation of hTERT promoter by EGF/Ets-2 was diminished via the EGFR kinase signal pathway-specific inhibitors AG1478 and Iressa. Inhibitors of Erk and Akt inhibited Ets-2-activated hTERT promoter activity. These data suggested that Ets-2, a genuine cancer-specific transcription factor, is actively involved in EGFR kinase-induced hTERT overexpression pathway in lung cancer cells. Blockage of this pathway may contribute to targeted gene therapy in lung cancer.

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References

  1. Vasa M, Breitschopf K, Zeiher AM, Dimmeler S. Nitric oxide activates telomerase and delays endothelial cell senescence. Circ Res. 2000;87:540–2.

    Article  CAS  PubMed  Google Scholar 

  2. Dwyer J, Li H, Xu D, Liu JP. Transcriptional regulation of telomerase activity: roles of the Ets transcription factor family. Ann N Y Acad Sci. 2007;1114:36–47.

    Article  CAS  PubMed  Google Scholar 

  3. Hsu CP, Miaw J, Hsia JY, Shai SE, Chen CY. Concordant expression of the telomerase-associated genes in non-small cell lung cancer. Eur J Surg Oncol. 2003;29:594–9.

    Article  PubMed  Google Scholar 

  4. Wu TC, Lin P, Hsu CP, Huang YJ, Chen CY, Chung WC, et al. Loss of telomerase activity may be a potential favorable prognostic marker in lung carcinomas. Lung Cancer. 2003;41:163–9.

    Article  PubMed  Google Scholar 

  5. Kyo S, Takakura M, Fujiwara T, Inoue M. Understanding and exploiting hTERT promoter regulation for diagnosis and treatment of human cancers. Cancer Sci. 2008;99:1528–38.

    Article  CAS  PubMed  Google Scholar 

  6. Cong YS, Wen J, Bacchetti S. The human telomerase catalytic subunit hTERT: organization of the gene and characterization of the promoter. Hum Mol Genet. 1999;8:137–42.

    Article  CAS  PubMed  Google Scholar 

  7. Hsu CP, Miaw J, Shai SE, Chen CY. Correlation between telomerase expression and terminal restriction fragment length ratio in non-small cell lung cancer—an adjusted measurement and its clinical significance. Eur J Cardiothorac Surg. 2004;26:425–31.

    Article  PubMed  Google Scholar 

  8. Wasylyk B, Wasylyk C, Flores P, Begue A, Leprince D, Stehelin D. The c-ets proto-oncogenes encode transcription factors that cooperate with c-Fos and c-Jun for transcriptional activation. Nature. 1990;346:191–3.

    Article  CAS  PubMed  Google Scholar 

  9. Borie R, Kannengiesser C, Crestani B. Familial forms of nonspecific interstitial pneumonia/idiopathic pulmonary fibrosis: clinical course and genetic background. Curr Opin Pulm Med. 2012;18:455–61.

    Article  PubMed  Google Scholar 

  10. Hsu CP, Hsu NY, Lee LW, Ko JL. Ets-2 binding site single nucleotide polymorphism at the hTERT gene promoter–effect on telomerase expression and telomere length maintenance in non-small cell lung cancer. Eur J Cancer. 2006;42:1466–74.

    Article  CAS  PubMed  Google Scholar 

  11. Park NH, Guo W, Kim HR, Kang MK. C-myc and sp1/3 are required for transactivation of hamster telomerase catalytic subunit gene promoter. Int J Oncol. 2001;19:755–61.

    CAS  PubMed  Google Scholar 

  12. Kim SJ, Lee HS, Shin JH, Kim CG, Jeong S, Park K, et al. Preferentially enhanced gene expression from a synthetic human telomerase reverse transcriptase promoter in human cancer cells. Oncol Rep. 2006;16:975–9.

    CAS  PubMed  Google Scholar 

  13. McMurray HR, McCance DJ. Human papillomavirus type 16 E6 activates TERT gene transcription through induction of c-Myc and release of USF-mediated repression. J Virol. 2003;77:9852–61.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Ito H, Kyo S, Kanaya T, Takakura M, Inoue M, Namiki M. Expression of human telomerase subunits and correlation with telomerase activity in urothelial cancer. Clin Cancer Res. 1998;4:1603–8.

    CAS  PubMed  Google Scholar 

  15. Liao CH, Hsiao YM, Hsu CP, Lin MY, Wang JC, Huang YL, et al. Transcriptionally mediated inhibition of telomerase of fungal immunomodulatory protein from ganoderma tsugae in A549 human lung adenocarcinoma cell line. Mol Carcinog. 2006;45:220–9.

    Article  CAS  PubMed  Google Scholar 

  16. Maida Y, Kyo S, Kanaya T, Wang Z, Yatabe N, Tanaka M, et al. Direct activation of telomerase by EGF through Ets-mediated transactivation of TERT via MAP kinase signaling pathway. Oncogene. 2002;21:4071–9.

    Article  CAS  PubMed  Google Scholar 

  17. Xiao X, Athanasiou M, Sidorov IA, Horikawa I, Cremona G, Blair D, et al. Role of Ets/Id proteins for telomerase regulation in human cancer cells. Exp Mol Pathol. 2003;75:238–47.

    Article  CAS  PubMed  Google Scholar 

  18. Dwyer JM, Liu JP. Ets-2 transcription factor, telomerase activity and breast cancer. Clin Exp Pharmacol Physiol. 2010;37:83–7.

    Article  CAS  PubMed  Google Scholar 

  19. Al-azawi D, Ilroy MM, Kelly G, Redmond AM, Bane FT, Cocchiglia S, et al. Ets-2 and p160 proteins collaborate to regulate c-myc in endocrine resistant breast cancer. Oncogene. 2008;27:3021–31.

    Article  CAS  PubMed  Google Scholar 

  20. Xu D, Dwyer J, Li H, Duan W, Liu JP. Ets-2 maintains hTERT gene expression and breast cancer cell proliferation by interacting with c-Myc. J Biol Chem. 2008;283:23567–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Xu D, Popov N, Hou M, Wang Q, Bjorkholm M, Gruber A, et al. Switch from Myc/Max to Mad1/Max binding and decrease in histone acetylation at the telomerase reverse transcriptase promoter during differentiation of HL60 cells. Proc Natl Acad Sci U S A. 2001;98:3826–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. McMahon SB, Wood MA, Cole MD. The essential cofactor TRRAP recruits the histone acetyltransferase hGCN5 to c-Myc. Mol Cell Biol. 2000;20:556–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Li CT, Hsiao YM, Wu TC, Lin YW, Yeh KT, Ko JL. Vorinostat, SAHA, represses telomerase activity via epigenetic regulation of telomerase reverse transcriptase in non-small cell lung cancer cells. J Cell Biochem. 2011;112:3044–53.

    Article  CAS  PubMed  Google Scholar 

  24. Tanno S, Ohsaki Y, Nakanishi K, Toyoshima E, Kikuchi K. Small cell lung cancer cells express EGFR and tyrosine phosphorylation of egfr is inhibited by gefitinib (“Iressa”, ZD1839). Oncol Rep. 2004;12:1053–7.

    CAS  PubMed  Google Scholar 

  25. Deng WG, Jayachandran G, Wu G, Xu K, Roth JA, Ji L. Tumor-specific activation of human telomerase reverses transcriptase promoter activity by activating enhancer-binding protein-2beta in human lung cancer cells. J Biol Chem. 2007;282:26460–70.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by the National Science Council, Taiwan, R.O.C. (NSC-97-2314-B-075-A-005-MY2). Real-time PCR was performed by the Instrument Center of Chung Shan Medical University, which is partly supported by the National Science Council, Ministry of Education, and Chung Shan Medical University.

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Authors and Affiliations

  1. Division of Thoracic Surgery, Department of Surgery, Taichung Veterans General Hospital, 160, Sec.3, Taichung-Kang Rd., Taichung, Taiwan

    Chung-Ping Hsu & Li-Wen Lee

  2. School of Medicine, National Yang-Ming University, Taipei, Taiwan

    Chung-Ping Hsu

  3. Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan

    Sheau-Chung Tang, I-Lun Hsin, Yu-Wen Lin & Jiunn-Liang Ko

  4. Departments of Chest Medicine and Medical Oncology, Chung Shan Medical University Hospital, Taichung, Taiwan

    Sheau-Chung Tang, I-Lun Hsin, Yu-Wen Lin & Jiunn-Liang Ko

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  1. Chung-Ping Hsu
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  2. Li-Wen Lee
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  3. Sheau-Chung Tang
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  4. I-Lun Hsin
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  5. Yu-Wen Lin
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  6. Jiunn-Liang Ko
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Correspondence to Chung-Ping Hsu or Jiunn-Liang Ko.

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Hsu, CP., Lee, LW., Tang, SC. et al. Epidermal growth factor activates telomerase activity by direct binding of Ets-2 to hTERT promoter in lung cancer cells. Tumor Biol. 36, 5389–5398 (2015). https://doi.org/10.1007/s13277-015-3204-x

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  • DOI: https://doi.org/10.1007/s13277-015-3204-x

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