hTERT phosphorylation by PKC is essential for telomerase holoprotein integrity and enzyme activity in head neck cancer cells

J. T. Chang, Y. C. Lu, Y. J. Chen, C. P. Tseng, Y. L. Chen, C. W. Fang, A. J. Cheng

Research output: Contribution to journalArticle

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Abstract

Telomerase activity is suppressed in normal somatic tissues but is activated in most cancer cells. We have previously found that all six telomerase subunit proteins, including hTERT and hsp90 are needed for full enzyme activity. Telomerase activity has been reported to be upregulated by protein kinase C (PKC), but the mechanism is not clear. In this study, we examined how PKC regulates telomerase activity in head and neck cancer cells. PKC inhibitor, bisindolylmaleimide I (BIS), inhibited telomerase activity but had no effect on the expressions of telomerase core subunits. RNA interference (RNAi) and in vitro phosphorylation studies revealed that PKC isoforms α, β, δ, ε, ζ specifically involved in telomerase regulation, and the phosphorylation target was on hTERT. Treatment with the hsp-90 inhibitor novobiocin dissociated hsp90 and hTERT as revealed by immunoprecipitation and immunoblot analysis and reduced telomerase activity. Treatment with the PKC activator SC-10 restored the association of hsp90 and hTERT and reactivate telomerase, suggesting that hTERT phosphorylation by PKC is essential for telomerase holoenzyme integrity and function. Analysis on clinical normal and tumour tissues reveal that the expressions of PKC α, β, δ, ε, ζ were higher in the tumour tissues, correlated with telomerase activity. Disruption of PKC phosphorylation by BIS significantly increased chemosensitivity to cisplatin. In conclusion, PKC isoenzyrnes α, β, δ, ε, ζ regulate telomerase activity in head and neck cancer cells by phosphorylating hTERT. This phosphorylation is essential for telomerase holoenzyme assembly, leading to telomerase activation and oncogenesis. Manipulation of telomerase activity by PKC inhibitors is worth exploring as an adjuvant therapeutic approach.

Original languageEnglish
Pages (from-to)870-878
Number of pages9
JournalBritish Journal of Cancer
Volume94
Issue number6
DOIs
Publication statusPublished - Mar 27 2006
Externally publishedYes

Fingerprint

Telomerase
Head and Neck Neoplasms
Protein Kinase C
Phosphorylation
Enzymes
Holoenzymes
Protein C Inhibitor
Protein Kinase Inhibitors
Novobiocin
Neoplasms
Protein Subunits
RNA Interference
Immunoprecipitation
Cisplatin

Keywords

  • hTERT
  • Phosphorylation
  • PKC isoenzymes
  • Telomerase

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

hTERT phosphorylation by PKC is essential for telomerase holoprotein integrity and enzyme activity in head neck cancer cells. / Chang, J. T.; Lu, Y. C.; Chen, Y. J.; Tseng, C. P.; Chen, Y. L.; Fang, C. W.; Cheng, A. J.

In: British Journal of Cancer, Vol. 94, No. 6, 27.03.2006, p. 870-878.

Research output: Contribution to journalArticle

Chang, J. T. ; Lu, Y. C. ; Chen, Y. J. ; Tseng, C. P. ; Chen, Y. L. ; Fang, C. W. ; Cheng, A. J. / hTERT phosphorylation by PKC is essential for telomerase holoprotein integrity and enzyme activity in head neck cancer cells. In: British Journal of Cancer. 2006 ; Vol. 94, No. 6. pp. 870-878.
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AB - Telomerase activity is suppressed in normal somatic tissues but is activated in most cancer cells. We have previously found that all six telomerase subunit proteins, including hTERT and hsp90 are needed for full enzyme activity. Telomerase activity has been reported to be upregulated by protein kinase C (PKC), but the mechanism is not clear. In this study, we examined how PKC regulates telomerase activity in head and neck cancer cells. PKC inhibitor, bisindolylmaleimide I (BIS), inhibited telomerase activity but had no effect on the expressions of telomerase core subunits. RNA interference (RNAi) and in vitro phosphorylation studies revealed that PKC isoforms α, β, δ, ε, ζ specifically involved in telomerase regulation, and the phosphorylation target was on hTERT. Treatment with the hsp-90 inhibitor novobiocin dissociated hsp90 and hTERT as revealed by immunoprecipitation and immunoblot analysis and reduced telomerase activity. Treatment with the PKC activator SC-10 restored the association of hsp90 and hTERT and reactivate telomerase, suggesting that hTERT phosphorylation by PKC is essential for telomerase holoenzyme integrity and function. Analysis on clinical normal and tumour tissues reveal that the expressions of PKC α, β, δ, ε, ζ were higher in the tumour tissues, correlated with telomerase activity. Disruption of PKC phosphorylation by BIS significantly increased chemosensitivity to cisplatin. In conclusion, PKC isoenzyrnes α, β, δ, ε, ζ regulate telomerase activity in head and neck cancer cells by phosphorylating hTERT. This phosphorylation is essential for telomerase holoenzyme assembly, leading to telomerase activation and oncogenesis. Manipulation of telomerase activity by PKC inhibitors is worth exploring as an adjuvant therapeutic approach.

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