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Apoptosis Versus Cell Death

CREB antisense oligonucleotides induce non-apoptotic cell death in proliferating leukemia cells, but not normal hematopoietic cells, by a bizarre non-antisense mechanism

Leukemia volume 15, pages 238–245 (2001)Cite this article

Abstract

We report that antisense phosphorothioate oligodeoxyribonucleotides (PS-ODNs) against cyclic AMP response element-binding protein (CREB) induce the death of human leukemia cell lines including HL-60, Kasumi-1 and K562, OCI-AML1a and also primary leukemia cells isolated from patients with acute myelocytic leukemia and chronic myelocytic leukemia in blastic crisis. In contrast, normal human bone marrow CD34+ cells and normal peripheral blood lymphocytes were resistant to the antisense-mediated cell death. We found that antisense-treated HL-60 cells had prominent nuclear fragmentations but lacked apoptotic features including internucleosomal DNA cleavage and TUNEL positivity. Cell cycle analysis demonstrated a remarkable reduction in G1 phase population along with a mild accumulation of S phase and good preservation of G2/M phase, indicating cells died at G2/M without cycling into G1 phase. None of the sense-sequenced PS-ODNs induced cell death. Further, neither the expression nor the message of CREB protein was reduced by antisense treatment, indicating that cell death was mediated by a non-antisense mechanism. On the other hand, no consensus oligonucleotide sequence for cell death induction was detected. Rather, we found a good correlation between the melting temperatures and the anti-proliferative activities of the oligonucleotides. Thus, CREB antisense PS-ODNs selectively induce a non-apoptotic cell death in leukemic cells by an unknown hybridization-dependent mechanism.

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Acknowledgements

This work was partially supported by a Grant-in-Aid for the Second Term Comprehensive 10-year Strategy for Cancer Control from the Ministry of Health and Welfare, Japan. We would like to thank Professor Nobuo Nara at Tokyo Medical and Dental University for providing human leukemia cell line OCI-AML1a.

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Authors and Affiliations

  1. Department of Hematology, Research Institute, International Medical Center of Japan, Tokyo, Japan

    K Saeki, A Yuo, F Takaku & Y Yazaki

  2. Exploratory Chemistry Research Laboratories, Sankyo Co. Ltd, Tokyo, Japan

    M Koizumi & M Kaneko

  3. Biological Research Laboratories, Sankyo Co. Ltd, Tokyo, Japan

    K Fujiwara

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  1. K Saeki
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Saeki, K., Yuo, A., Koizumi, M. et al. CREB antisense oligonucleotides induce non-apoptotic cell death in proliferating leukemia cells, but not normal hematopoietic cells, by a bizarre non-antisense mechanism. Leukemia 15, 238–245 (2001). https://doi.org/10.1038/sj.leu.2402014

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