Long-term suppression of telomerase expression in HeLa cell clones, transfected with an expression vector carrying siRNA targeting hTERT mRNA

  • Authors:
    • Kaisa Kurvinen
    • Stina Syrjänen
    • Bo Johansson
  • View Affiliations

  • Published online on: July 1, 2006     https://doi.org/10.3892/ijo.29.1.279
  • Pages: 279-288
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Abstract

HeLa cell cultures were used as model systems for small interfering RNA (siRNA) induced knockdown of mRNA expression of the human telomerase catalytic subunit, telomerase reverse transcriptase (hTERT). Four 21-bp siRNAs targeting different sites of the hTERT mRNA were designed, and the siRNA molecules produced by a T7 transcription system in vitro. In transient transfection assays on HeLa cells, only one of the tested siRNAs produced a potent knockdown effect on hTERT mRNA expression, associated with the suppression of telomerase activity (both reduced by ≈50%). An expression vector encoding a hairpin siRNA against the effective hTERT mRNA target site was generated, and HeLa clones stably expressing hTERT-specific siRNA were created. Two clones showed extremely reduced hTERT mRNA expression, associated with unusually short telomeres, the inhibition of cell growth and the induction of senescence and apoptosis. Thus, there was obvious loss of viability in cells lacking hTERT expression and carrying short telomeres. This was most prominent in the clone that showed prolonged reductions (over two months) in both hTERT expression and telomerase activity. Thus, our data clearly show that long-term suppression of telomerase expression by siRNA is an attainable goal, at least in a HeLa cell model system.

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July 2006
Volume 29 Issue 1

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Kurvinen K, Syrjänen S and Johansson B: Long-term suppression of telomerase expression in HeLa cell clones, transfected with an expression vector carrying siRNA targeting hTERT mRNA. Int J Oncol 29: 279-288, 2006
APA
Kurvinen, K., Syrjänen, S., & Johansson, B. (2006). Long-term suppression of telomerase expression in HeLa cell clones, transfected with an expression vector carrying siRNA targeting hTERT mRNA. International Journal of Oncology, 29, 279-288. https://doi.org/10.3892/ijo.29.1.279
MLA
Kurvinen, K., Syrjänen, S., Johansson, B."Long-term suppression of telomerase expression in HeLa cell clones, transfected with an expression vector carrying siRNA targeting hTERT mRNA". International Journal of Oncology 29.1 (2006): 279-288.
Chicago
Kurvinen, K., Syrjänen, S., Johansson, B."Long-term suppression of telomerase expression in HeLa cell clones, transfected with an expression vector carrying siRNA targeting hTERT mRNA". International Journal of Oncology 29, no. 1 (2006): 279-288. https://doi.org/10.3892/ijo.29.1.279