Abstract
We have evaluated the apoptotic and DNA damaging activity of Idarubicin (IDA) on K-562 cells alone and following the uptake of modified antisense-oligodeoxynucleotides (AS-ODNs) targeting b3a2 mRNA of bcr/abl hybrid gene, after treatment with AS-ODNs/DCChol-DOPE (liposomes) complexes. The uptake of FITC-labeled oligonucleotide-liposomes complexes (FITC-ODNs/DCChol-DOPE) was analyzed by flow cytometry and fluorescence microscopy. Both techniques indicated cytoplasmic accumulation of labeled liposome complexes following 24h of exposure. In absence of liposomes, AS-ODNs uptake was minimal. Pre-treatment of cells with AS-ODNs/DCChol-DOPE increased the capability of IDA to induce apoptosis as determined by morphology and the comet assay. In contrast, the use of a non-sense oligodeoxynucleotide conjugated with liposomes, in the presence of IDA, did not increase K-562 cell apoptosis. Nevertheless, DNA damage in IDA treated cells was not related to ODNs/liposomes pre-treatment, as determined by the comet assay. Our data suggests that DCChol-DOPE increases the uptake of ODNs in K-562 cells, and these modified AS-ODNs increase IDA induced apoptosis by decreasing p210(bcr/abl) levels in K-562 cells.
MeSH terms
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Apoptosis / drug effects*
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Biological Transport
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Cations / chemistry
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Cholesterol / analogs & derivatives*
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Cholesterol / chemistry
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Comet Assay
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DNA Damage
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DNA, Neoplasm / analysis
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Drug Synergism
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Flow Cytometry
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Fusion Proteins, bcr-abl / biosynthesis
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Fusion Proteins, bcr-abl / genetics
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Gene Expression Regulation, Leukemic / drug effects
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Humans
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Idarubicin / pharmacology*
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K562 Cells / drug effects*
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K562 Cells / metabolism
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Lipids / chemistry
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Liposomes / administration & dosage
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Liposomes / chemistry
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Microscopy, Fluorescence
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Neoplasm Proteins / biosynthesis
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Neoplasm Proteins / genetics
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Oligodeoxyribonucleotides, Antisense / administration & dosage
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Oligodeoxyribonucleotides, Antisense / metabolism*
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Oligodeoxyribonucleotides, Antisense / pharmacology
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Phosphatidylethanolamines / chemistry
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Quaternary Ammonium Compounds / chemistry
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RNA, Messenger / antagonists & inhibitors
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RNA, Messenger / genetics
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RNA, Neoplasm / antagonists & inhibitors
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RNA, Neoplasm / genetics
Substances
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Cations
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DNA, Neoplasm
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Lipids
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Liposomes
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Neoplasm Proteins
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Oligodeoxyribonucleotides, Antisense
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Phosphatidylethanolamines
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Quaternary Ammonium Compounds
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RNA, Messenger
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RNA, Neoplasm
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3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol
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(3-dimyristyloxypropyl)(dimethyl)(hydroxyethyl)ammonium
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dioleoyl phosphatidylethanolamine
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Cholesterol
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Fusion Proteins, bcr-abl
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Idarubicin