TY - JOUR
T1 - Antisense BCR-ABL oligonucleotides induce apoptosis in the philadelphia chromosome-positive cell line BV173
AU - Smetsers, Toon F.C.M.
AU - Skorski, Tomasz
AU - Van De Locht, Louis T.F.
AU - Wessels, Hans M.C.
AU - Pennings, Arie H.M.
AU - De Witte, Theo
AU - Calabretta, Bruno
AU - Mensink, Ewald J.B.M.
PY - 1994/1
Y1 - 1994/1
N2 - BCR-ABL antisense oligonucleotides can specifically reduce colony formation of early hematopoietic progenitor cells from chronic myeloid leukemia (CML) patients. Little is known about the mechanism of this inhibition. We studied the inhibition of the bcr-abl oncogene using fluorescein-labeled phosphorothioate oligonucleotides in the Philadelphia chromosome-positive cell line BV173. Oligonucleotide stability, uptake, bcr-abl mRNA degradation, inhibition of cell proliferation, and cell death were studied. The oligonucleotide uptake was directly dependent on the extracellular concentration and was constant over the first 18h of incubation. After that the uptake rate decreased. We detected a decrease in bcr-abl mRNA after 3 days of treatment with antisense oligonucleotides, but much less in controls. The controls used in the experiments were the sense oligonucleotide, equimolar amounts of sense and antisense, and an untreated control. Antisense oligonucleotides completely inhibited cell growth of BV173 cells and did not inhibit growth of HL-60 cells, whereas control oligonucleotides had no such effect on either cell line. An oligonucleotide specific for the other CML breakpoint was also effective in reducing cell growth of BV173. By the use of a DNA double staining technique to discriminate between necrotic and apoptotic cells, we detected a large number of apoptotic cells in antisense treated BV173 cultures after 5 days of treatment as compared to controls. We conclude that antisense BCR-ABL oligonucleotides reduce bcr-abl mRNA expression in BV173 cells mainly in a sequence-specific manner and induce apoptosis.
AB - BCR-ABL antisense oligonucleotides can specifically reduce colony formation of early hematopoietic progenitor cells from chronic myeloid leukemia (CML) patients. Little is known about the mechanism of this inhibition. We studied the inhibition of the bcr-abl oncogene using fluorescein-labeled phosphorothioate oligonucleotides in the Philadelphia chromosome-positive cell line BV173. Oligonucleotide stability, uptake, bcr-abl mRNA degradation, inhibition of cell proliferation, and cell death were studied. The oligonucleotide uptake was directly dependent on the extracellular concentration and was constant over the first 18h of incubation. After that the uptake rate decreased. We detected a decrease in bcr-abl mRNA after 3 days of treatment with antisense oligonucleotides, but much less in controls. The controls used in the experiments were the sense oligonucleotide, equimolar amounts of sense and antisense, and an untreated control. Antisense oligonucleotides completely inhibited cell growth of BV173 cells and did not inhibit growth of HL-60 cells, whereas control oligonucleotides had no such effect on either cell line. An oligonucleotide specific for the other CML breakpoint was also effective in reducing cell growth of BV173. By the use of a DNA double staining technique to discriminate between necrotic and apoptotic cells, we detected a large number of apoptotic cells in antisense treated BV173 cultures after 5 days of treatment as compared to controls. We conclude that antisense BCR-ABL oligonucleotides reduce bcr-abl mRNA expression in BV173 cells mainly in a sequence-specific manner and induce apoptosis.
KW - Apoptosis/drug effects
KW - Base Sequence
KW - Cell Death
KW - Cell Division/drug effects
KW - DNA Damage
KW - Fluoresceins
KW - Fusion Proteins, bcr-abl/genetics
KW - Humans
KW - Kinetics
KW - Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy
KW - Molecular Sequence Data
KW - Oligonucleotides, Antisense/metabolism
KW - RNA, Messenger/genetics
KW - Sensitivity and Specificity
KW - Tumor Cells, Cultured
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UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=purepublist2023&SrcAuth=WosAPI&KeyUT=WOS:A1994MT22100022&DestLinkType=FullRecord&DestApp=WOS
M3 - Article
C2 - 8289478
SN - 0887-6924
VL - 8
SP - 129
EP - 140
JO - Leukemia
JF - Leukemia
IS - 1
ER -