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BCR-ABL antisense oligodeoxyribonucleotides suppress the growth of leukemic and normal hematopoietic cells by a
J L Vaerman1, C Lammineur, P Moureau
1Laboratoire de Biologie Moléculaire Clinique et Unité d'Hématologie, Cliniques St Luc, Université Catholique de Louvain, Bruxelles, Belgium.
Abstract:
We have examined the effect of BCR/ABL junctional antisense phosphodiester oligodeoxyribonucleotides (ODNs) on BV173 and other chronic myeloid leukemia (CML) cell lines. Various control ODNs were used to understand the mechanism of the observed antiproliferative effect. Not only the antisense ODNs but also several control ODNs inhibit the proliferation of the leukemic cell lines. All the ODNs that inhibit the cell proliferation share a TAT consensus sequence at their 3' end. A 1-base mismatch within this consensus sequence abolishes the antiproliferative effect. Mismatches of several bases at any other position within the sequence of the active ODNs do not suppress the observed effect. Similar experiments on normal or CML CD34+ cell fraction led to the same observations. We conclude that the antiproliferative effect of the phosphodiester BCR/ABL antisense ODNs cannot be attributed to an antisense mechanism but rather to a nonelucidated effect of a 3' terminal TAT sequence. This effect is not CML specific.
Insights
Antisense oligodeoxyribonucleotides (ODNs) targeting BCR/ABL showed antiproliferative effects in chronic myeloid leukemia (CML) cells. However, this effect was linked to a 3' TAT sequence, not an antisense mechanism.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Chronic myeloid leukemia (CML) is characterized by the BCR/ABL fusion gene.
- Antisense oligodeoxyribonucleotides (ODNs) are investigated as potential therapeutics for CML.
- The precise mechanism of ODN action in CML requires further elucidation.
Purpose of the Study:
- To investigate the antiproliferative effects of BCR/ABL antisense ODNs on CML cell lines.
- To determine the mechanism underlying the observed antiproliferative activity.
- To identify specific sequence requirements for ODN-mediated cell growth inhibition.
Main Methods:
- Treatment of CML cell lines (BV173) and CD34+ cells with various phosphodiester ODNs.
- Inclusion of control ODNs with modifications to elucidate the mechanism of action.
- Analysis of cell proliferation and sequence-structure activity relationships.
Main Results:
- Both BCR/ABL antisense ODNs and control ODNs exhibited antiproliferative effects on CML cell lines.
- A conserved 3' terminal TAT consensus sequence was critical for the antiproliferative activity.
- A single base mismatch within the TAT sequence abolished the effect, while other mismatches did not.
Conclusions:
- The antiproliferative effect of these ODNs is not mediated by an antisense mechanism.
- A non-elucidated mechanism involving a 3' terminal TAT sequence drives the observed inhibition.
- This TAT sequence-dependent effect is not specific to CML.