2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells

David M. Kushner, Jayashree M. Paranjape, Bhaswati Bandyopadhyay, Hagen Cramer, Douglas W. Leaman, Alexander W. Kennedy, Robert H. Silverman, John Kenneth Cowell

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Objective. RNase L is converted to an active form upon binding short 2',5'-oligoadenylates (2-5A). To direct RNase L to an RNA target, 2-5A is attached to an antisense oligonucleotide (2-5A antisense). This chimera can be directed against telomerase-an RNA-protein complex that elongates telomeric DNA and is involved in cellular immortalization. Our objective is to investigate the effect of 2-5A antisense by targeting telomerase RNA (hTR) in the ovarian cancer cell line, HEY-1B. Methods. Baseline RNase L levels and telomerase activities were measured in both HEY-1B and normal ovarian epithelial cells (NOE). Cells were treated daily with chimeric oligonucleotides (ODN) directed against four different hTR sites, or control ODNs including nonchimeric antisense, 2-5A fused to a mismatched sequence, or inactive 2-5A fused to antisense. At 48 h, apoptosis was evaluated using the TUNEL assay. After six daily ODN administrations, telomerase activity was redetermined, and at 7 days viability counts were obtained. Results. Both cell lines expressed similar levels of RNase L. Hey-1B displayed telomerase activity while NOE did not. After 7 days of transfection, 2-5A antisense ODNs caused profound cell death in the HEY-1B cells, but not in the NOE cells. This effect was seen regardless of hTR target site, and ODN controls showed no significant decrease in cell viability in either cell line. HEY1B cells treated with 2-5A antisense against hTR showed a decrease in telomerase activity and a profound induction of programmed cell death. Conclusions. The results suggest that 2-5A antisense directed against telomerase RNA results in apoptotic cell death in ovarian cancer cells, but not normal ovarian epithelial cells. The 2-5A antisense strategy may hold a considerable advantage over the conventional antisense approach in targeting cancer- causing genes.

Original languageEnglish (US)
Pages (from-to)183-192
Number of pages10
JournalGynecologic Oncology
Volume76
Issue number2
DOIs
StatePublished - Jan 1 2000
Externally publishedYes

Fingerprint

Ovarian Neoplasms
Apoptosis
Telomerase
Epithelial Cells
Cell Death
Cell Line
telomerase RNA
2',5'-oligoadenylate
Antisense Oligonucleotides
Neoplasm Genes
In Situ Nick-End Labeling
Oligonucleotides
Transfection
Cell Survival
RNA
2-5A-dependent ribonuclease
DNA

Keywords

  • 2-5A
  • Antisense
  • Ovarian cancer
  • Telomerase

ASJC Scopus subject areas

  • Oncology
  • Obstetrics and Gynecology

Cite this

Kushner, D. M., Paranjape, J. M., Bandyopadhyay, B., Cramer, H., Leaman, D. W., Kennedy, A. W., ... Cowell, J. K. (2000). 2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells. Gynecologic Oncology, 76(2), 183-192. https://doi.org/10.1006/gyno.1999.5668

2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells. / Kushner, David M.; Paranjape, Jayashree M.; Bandyopadhyay, Bhaswati; Cramer, Hagen; Leaman, Douglas W.; Kennedy, Alexander W.; Silverman, Robert H.; Cowell, John Kenneth.

In: Gynecologic Oncology, Vol. 76, No. 2, 01.01.2000, p. 183-192.

Research output: Contribution to journalArticle

Kushner, DM, Paranjape, JM, Bandyopadhyay, B, Cramer, H, Leaman, DW, Kennedy, AW, Silverman, RH & Cowell, JK 2000, '2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells', Gynecologic Oncology, vol. 76, no. 2, pp. 183-192. https://doi.org/10.1006/gyno.1999.5668
Kushner DM, Paranjape JM, Bandyopadhyay B, Cramer H, Leaman DW, Kennedy AW et al. 2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells. Gynecologic Oncology. 2000 Jan 1;76(2):183-192. https://doi.org/10.1006/gyno.1999.5668
Kushner, David M. ; Paranjape, Jayashree M. ; Bandyopadhyay, Bhaswati ; Cramer, Hagen ; Leaman, Douglas W. ; Kennedy, Alexander W. ; Silverman, Robert H. ; Cowell, John Kenneth. / 2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells. In: Gynecologic Oncology. 2000 ; Vol. 76, No. 2. pp. 183-192.
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abstract = "Objective. RNase L is converted to an active form upon binding short 2',5'-oligoadenylates (2-5A). To direct RNase L to an RNA target, 2-5A is attached to an antisense oligonucleotide (2-5A antisense). This chimera can be directed against telomerase-an RNA-protein complex that elongates telomeric DNA and is involved in cellular immortalization. Our objective is to investigate the effect of 2-5A antisense by targeting telomerase RNA (hTR) in the ovarian cancer cell line, HEY-1B. Methods. Baseline RNase L levels and telomerase activities were measured in both HEY-1B and normal ovarian epithelial cells (NOE). Cells were treated daily with chimeric oligonucleotides (ODN) directed against four different hTR sites, or control ODNs including nonchimeric antisense, 2-5A fused to a mismatched sequence, or inactive 2-5A fused to antisense. At 48 h, apoptosis was evaluated using the TUNEL assay. After six daily ODN administrations, telomerase activity was redetermined, and at 7 days viability counts were obtained. Results. Both cell lines expressed similar levels of RNase L. Hey-1B displayed telomerase activity while NOE did not. After 7 days of transfection, 2-5A antisense ODNs caused profound cell death in the HEY-1B cells, but not in the NOE cells. This effect was seen regardless of hTR target site, and ODN controls showed no significant decrease in cell viability in either cell line. HEY1B cells treated with 2-5A antisense against hTR showed a decrease in telomerase activity and a profound induction of programmed cell death. Conclusions. The results suggest that 2-5A antisense directed against telomerase RNA results in apoptotic cell death in ovarian cancer cells, but not normal ovarian epithelial cells. The 2-5A antisense strategy may hold a considerable advantage over the conventional antisense approach in targeting cancer- causing genes.",
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AU - Paranjape, Jayashree M.

AU - Bandyopadhyay, Bhaswati

AU - Cramer, Hagen

AU - Leaman, Douglas W.

AU - Kennedy, Alexander W.

AU - Silverman, Robert H.

AU - Cowell, John Kenneth

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