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  • Original Paper
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Killing of leukemic cells with a BCR/ABL fusion gene by RNA interference (RNAi)

Oncogene volume 21, pages 5716–5724 (2002)Cite this article

Abstract

Short 21-mer double-stranded RNA (dsRNA) molecules have recently been employed for the sequence-specific silencing of endogenous human genes. This mechanism, called RNA interference (RNAi), is extremely potent and requires only a few dsRNA molecules per cell to silence homologous gene mRNA expression. We used dsRNA targeting the M-BCR/ABL fusion site to kill leukemic cells with such a rearrangement. Transfection of dsRNA specific for the M-BCR/ABL fusion mRNA into K562 cells depleted the corresponding mRNA and the M-BCR/ABL oncoprotein. This was demonstrated by real-time quantitative PCR and Western blots. The BCR/ABL knockdown was accompanied by strong induction of apoptotic cell death. Leukemic cells without BCR/ABL rearrangement were not killed by M-BCR/ABL-dsRNA. In addition, to corroborate the extraordinary sequence specificity of RNAi, we designed another RNA oligo matching the M-BCR/ABL fusion site but having two point mutations within its central region. We show that these two point mutations abolished both p210 reduction and induction of apoptosis in K562 cells. Finally, we compared leukemic cell killing by RNAi to that caused by the ABL kinase tyrosine inhibitor, STI 571, Imatinib. For full induction of apoptosis, dsRNA targeting M-BCR/ABL required 24 h more than Imatinib. This may be caused by the relatively long half-life of the BCR/ABL oncoprotein, which is not targeted by the RNAi mechanism, but is affected by STI 571. When we applied ds M-BCR/ABL RNA and STI 571 in combination, we did not observe a further increase in the induction of apoptosis. Nevertheless, these data may open a field for further studies towards gene-therapeutic approaches using RNA interference to kill tumor cells with specific genetic abnormalities.

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Acknowledgements

The authors wish to thank Dr T Tuschl, Dr S Viehmann and D Rawer for their help with the design of the dsRNA, quantification of BCR/ABL mRNA by taqman PCR, or generation of plasmid standards, respectively. Expert technical assistance by Stefanie Garkisch is gratefully acknowledged. A part of these studies was supported by a grant from the Deutsche Forschungsgemeinschaft to A Borkhardt.

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  1. Monika Wilda, Uta Fuchs, Wilhelm Wössmann and Arndt Borkhardt: These authors contributed equally to this work

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  1. Department of Pediatric Hematology and Oncology, University of Giessen, Giessen, 35392, Germany

    Monika Wilda, Uta Fuchs, Wilhelm Wössmann & Arndt Borkhardt

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  1. Monika Wilda
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Correspondence to Arndt Borkhardt.

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Wilda, M., Fuchs, U., Wössmann, W. et al. Killing of leukemic cells with a BCR/ABL fusion gene by RNA interference (RNAi). Oncogene 21, 5716–5724 (2002). https://doi.org/10.1038/sj.onc.1205653

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