Abstract
We compared antisense phosphorothioate oligonucleotides (PS-ODN) that target BCL-2 such as Genasense (G3139-PS), with other PS-ODN or phosphodiester-ODN (PO-ODN) in their relative capacity to induce apoptosis of chronic lymphocytic leukemia (CLL) B cells in vitro. Surprisingly, we found that thymidine-containing PS-ODN, but not PO-ODN, induced activation and apoptosis of CLL cells independent of BCL-2 antisense sequence or CpG motifs. All tested thimidine-containing PS-ODN, irrespective of their primary sequences, reduced the expression of Bcl-2 protein and increased the levels of the proapoptotic molecules p53, Bid, Bax in CLL cells. Apoptosis induced by thymidine-containing PS-ODN was preceded by cellular activation, could be blocked by the tyrosine-kinase inhibitor imatinib mesylate (Gleevec), and was dependent on ABL kinase. We conclude that thymidine-containing PS-ODN can activate CLL cells and induce apoptosis via a mechanism that is independent of BCL-2 gene interference or CpG motifs.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Chloromethyl Ketones / pharmacology
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Apoptosis / drug effects
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Apoptosis / physiology
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B-Lymphocytes / drug effects*
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BH3 Interacting Domain Death Agonist Protein / drug effects
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BH3 Interacting Domain Death Agonist Protein / metabolism
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Benzamides
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Caspases / drug effects
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Caspases / metabolism
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Cell Survival / drug effects
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CpG Islands / drug effects
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CpG Islands / genetics*
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CpG Islands / physiology
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Drug Screening Assays, Antitumor
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Enzyme Activation / drug effects
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Genes, bcl-2 / drug effects
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Genes, bcl-2 / genetics*
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Genes, bcl-2 / physiology
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Humans
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Imatinib Mesylate
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In Vitro Techniques
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Leukemia, Lymphocytic, Chronic, B-Cell / metabolism*
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Oligodeoxyribonucleotides / antagonists & inhibitors
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Oligodeoxyribonucleotides / chemistry
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Oligodeoxyribonucleotides / pharmacology*
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Organothiophosphorus Compounds / chemistry
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Organothiophosphorus Compounds / pharmacology*
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Phosphorylation
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Piperazines / pharmacology
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Proto-Oncogene Proteins c-abl / drug effects
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Proto-Oncogene Proteins c-abl / metabolism
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Pyrimidines / pharmacology
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Structure-Activity Relationship
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Thymidine / chemistry*
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Thymidine / pharmacology
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Tumor Cells, Cultured
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Tumor Suppressor Protein p53 / drug effects
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Tumor Suppressor Protein p53 / metabolism
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Up-Regulation
Substances
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Amino Acid Chloromethyl Ketones
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BH3 Interacting Domain Death Agonist Protein
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BID protein, human
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Benzamides
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Oligodeoxyribonucleotides
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Organothiophosphorus Compounds
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Piperazines
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Pyrimidines
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Tumor Suppressor Protein p53
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benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
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Imatinib Mesylate
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Proto-Oncogene Proteins c-abl
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Caspases
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Thymidine