Enhanced uptake of antisense oligonucleotides using cationic liposomes and the apoptotic effect of idarubicin in K-562 cell line.
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.
Vellón
Luciano
L
Departamento de Genetica, Instituto de Investigaciones Hematológicas Mariano R. Castex, Academia Nacional de Medicina, J.A. Pacheco de Melo 3081, 1425, Buenos Aires, Argentina. lvellon@hematologia.anm.edu.ar
González-Cid
Marcela
M
Karara
Armando
A
Nebel
Marcelo de Campos
Mde C
Cuello
María Teresa
MT
Larripa
Irene
I
D017209:Apoptosis / Q000187:drug effects*; D001692:Biological Transport; D002412:Cations / Q000737:chemistry; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry; D020552:Comet Assay; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000032:analysis; D004357:Drug Synergism; D005434:Flow Cytometry; D016044:Fusion Proteins, bcr-abl / Q000096:biosynthesis / Q000235:genetics; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006801:Humans; D015255:Idarubicin / Q000494:pharmacology*; D020014:K562 Cells / Q000187:drug effects* / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000378:metabolism* / Q000494:pharmacology; D010714:Phosphatidylethanolamines / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics; D012334:RNA, Neoplasm / Q000037:antagonists & inhibitors / Q000235:genetics
D002412:Cations; D004273:DNA, Neoplasm; D008055:Lipids; D008081:Liposomes; D009363:Neoplasm Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D012334:RNA, Neoplasm; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; C087276:(3-dimyristyloxypropyl)(dimethyl)(hydroxyethyl)ammonium; C094877:dioleoyl phosphatidylethanolamine; D002784:Cholesterol; D016044:Fusion Proteins, bcr-abl; D015255:Idarubicin