Inhibition of Telomerase Activity of Melanoma Cellsin Vitroby Antisense Oligonucleotides

Глухов, А.И.; Zimnik, O.V.; Гордеев, С. А.; Северин, С. Е.

doi:10.1006/bbrc.1998.8801

Cited by 86 publications

(46 citation statements)

References 28 publications

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“…Following this appealing scienti®c rationale, several classes of telomerase inhibitors were prepared and evaluated. Among the compounds tested were small molecules (Naasani et al, 1998(Naasani et al, , 1999Bare et al, 1998;Hisatake et al, 1999), compounds capable of interacting with DNA G-quadruplex structures (Neidle et al, 2000;Hurley et al, 2000), dominant negative hTERTderived proteins (Hahn et al, 1999), antisense RNA (Feng et al, 1995;Yamaguchi et al, 1999), and various types of oligonucleotides, including DNA phosphodiesters (Glukhov et al, 1998), 2-5A tethered phosphodiesters (Kondo et al, 1998), phosphorothioates (Norton et al, 1996;Matthes and Lehmann, 1999;Sharma et al, 1996), phosphoramidates (Gryaznov et al, 2001), 2'-O-Methyl and 2'-O-(Methoxy-ethyl) phosphorothioate chimera (Pitts and Corey, 1998;Elayadi et al, 2001), ribozymes (Wan et al, 1998), and PNA molecules (Norton et al, 1996). The e ects of DNA, PNA and 2'-modi®ed RNA oligonucleotides reportedly were sequence speci®c, unlike the activity of the majority of earlier generation of phosphorothioates oligomers (Matthes and Lehman, 1999).…”

Section: Introductionmentioning

confidence: 99%

Oligonucleotide N3′→P5′ phosphoramidates as efficient telomerase inhibitors

et al. 2002

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Human telomerase is a unique reverse transcriptase that is expressed in multiple cancers, but not in the vast majority of normal cells. The enzyme is responsible for telomere protection and maintenance, and supports the proliferative immortality of cancer cells. Thus, it has been proposed that the speci®c inhibition of telomerase activity in tumors might have signi®cant and bene®cial therapeutic e ects. To this goal we have designed, synthesized, and evaluated several oligonucleotide N3'?P5' phosphoramidates as telomerase inhibitors. These oligonucleotides are complementary to the template region of the RNA domain of telomerase (hTR). The prepared compounds were evaluated in HME50-5E breast epithelial cells, where their e ects on telomerase activity were determined using a cell-based telomerase (TRAP) assay at 24 as well as 72 h after exposure to compounds. The oligo-N3'?P5' phosphoramidate inhibited telomerase activity in cells in the presence of the cellular up-take enhancer (FuGENE6 TM ) in a dose-and sequence-dependent manner, with IC 50 values of approximately 1 nM. Inhibition of telomerase activity by this compound without the lipid carrier was not e cient. However, the isosequential oligonucleotide N3'?P5' thio-phosphoramidate was able to inhibit telomerase activity with or without lipid carriers at nM, or low-mM concentrations, respectively. This inhibition of telomerase activity in HME50-5E cells by the oligonucleotide thio-phosphoramidates was also sequence speci®c. Long-term treatment of the cells with 0.5 mM of FuGENE6 formulated 13-mer thio-phosphoramidates, fully complementary to hTR, resulted in gradual telomere shortening, followed by cellular senescence and apoptosis, as would be predicted for a telomerase inhibitor. The mismatched control compound had no e ect on cell proliferation. The results suggest that the oligonucleotide N3'?P5' phosphoramidates, and particularly thio-phosphoramidates, might be further developed as selective anti-telomerase reagents.

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Section: Introductionmentioning

confidence: 99%

Oligonucleotide N3′→P5′ phosphoramidates as efficient telomerase inhibitors

et al. 2002

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“…15,26 Therefore, it is expected that inhibition of telomerase Gene Therapy function may alter the growth properties of tumor cells and subsequently suppress the tumor growth. Since the RNA component of human telomerase (hTR) was first of all reported, 22 several strategies that target hTR with antisense RNA, [22][23][24] hammer head ribozyme 33 or peptide nucleic acids 34 were investigated. They reduced telomerase activity and tumor growth to some extent.…”

Section: Discussionmentioning

confidence: 99%

“…These facts strongly support the idea that telomerase inhibition may represent a novel targeting approach for the treatment of malignant gliomas. Recently, it has been demonstrated that targeting the RNA template of telomerase (hTR) with antisense RNA [22][23][24][25] or inhibiting the function of a catalytic protein subunit of telomerase (hTERT) with a dominant-negative mutant [26][27][28] showed a significant antitumor effect. We have recently synthesized a 19-mer antisense oligonucleotide against hTR linked 2-5A (5′-phosphorylated 2′-5′-linked oligoadenylate) (2-5A-anti-hTR) and demonstrated its cytotoxic effect on malignant glioma cells 29,30 or prostate cancer cells.…”

Section: Introductionmentioning

confidence: 99%

Combination therapy of malignant glioma cells with 2-5A-antisense telomerase RNA and recombinant adenovirus p53

et al. 2000

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“…23,24 Since the RNA component of human telomerase (hTR) was first known, 25 several trials that target hTR with antisense RNA, [25][26][27] hammerhead ribozyme 28 or peptide nucleic acids 29 were reported. They reduced telomerase activity and tumor growth to some extent.…”

Section: Oligonucleotide Mixed With Lipofectamine (4 L/ml) Every 24mentioning

confidence: 99%

Treatment of bladder cancer cells in vitro and in vivo with 2–5A antisense telomerase RNA

et al. 2001

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Bladder cancer is the most common malignant tumor of the urinary tract. Novel treatment approaches are essential because of the failure of current treatment options to cure a high percentage of patients. Telomerase, a ribonucleoprotein, is detected in almost all bladder cancer, but not in normal bladder tissues. Therefore, telomerase is expected to be a very promising candidate for targeted therapy of bladder cancer. In this study, we synthesized a 19-mer antisense oligonucleotide against the RNA component of human telomerase (hTR) linked to a 2-5A molecule (2-5A-anti-hTR) and investigated its antitumor effect against bladder cancer cells. The 2-5A antisense strategy relies on the recruitment and activation of RNase L at the site of targeted RNA Keywords: 2-5A; antisense; telomerase; bladder cancer; apoptosisIn 1998 it was estimated that the number of new cases of bladder cancer would be greater than 54 000 and approximately 12 500 bladder cancer deaths would occur in the United States. 1 Superficial tumors including carcinoma in situ can be cured by transurethral resection and/or intravesical instillation of bacillus Calmette-Guérin vaccine (BCG). 2,3 However, 15% of these tumors progress to muscle-invasive cancer and required cystectomy. 4,5 In contrast with superficial tumors, the prognosis of advanced bladder cancer is poor and the 5-year survival rate is about 50% even after the radical surgical excision. 6 Therefore, in order to improve prognosis of bladder cancer, it is absolutely essential to explore a novel modality of treatment.Telomerase is a ribonucleoprotein enzyme that elongates telomeric DNA at the ends of chromosomes. 7 Most normal cells do not have the enzyme and lose telomeric DNA with each mitotic cycle, resulting in senescence or cell death. 7,8 By contrast, approximately 90% of tumors of wide range express telomerase and can continue to proliferate. [9][10][11] Recent investigations demonstrate that telomerase activity is detected in more than 90% of bladder cancer, but not in normal bladder tissues. 12,13 Therefore,

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Inhibition of Telomerase Activity of Melanoma Cellsin Vitroby Antisense Oligonucleotides

Cited by 86 publications

References 28 publications

Oligonucleotide N3′→P5′ phosphoramidates as efficient telomerase inhibitors

Oligonucleotide N3′→P5′ phosphoramidates as efficient telomerase inhibitors

Combination therapy of malignant glioma cells with 2-5A-antisense telomerase RNA and recombinant adenovirus p53

Treatment of bladder cancer cells in vitro and in vivo with 2–5A antisense telomerase RNA

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