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Chronic Myeloid Leukemia, BCR/ABL Studies and Myeloproliferative Disorders

Transient or long-term silencing of BCR-ABL alone induces cell cycle and proliferation arrest, apoptosis and differentiation

Leukemia volume 20, pages 68–76 (2006)Cite this article

Abstract

BCR-ABL fusion protein, a t(9;22) translocation product is indispensable for generation, maintenance and progression of chronic myeloid leukemia. RNA interference is an approach to silence gene at post-transcriptional level. We show that dsRNA targeted against the translocation region leads to more than 90% inhibition of BCR-ABL mRNA and protein expression levels using K562 as a model. Lack of BCR-ABL leads to cell cycle arrest in G1 phase as observed by decrease in cyclin D1 and increase in p21 and p27 cdk inhibitors mRNA. Apoptosis resistance imparted by BCR-ABL is lost in these cells in caspase-dependent or independent manner. Decrease in Bcl-XL is observed along with decrease in mitochondrial membrane integrity. Transient removal of BCR-ABL expression has a profound effect on proliferation and clonogenic capacity also confirmed by long-term silencing using lentiviral vectors. Interestingly, low level of BCR-ABL message leads to enhanced erythroid differentiation and reduced expression of megakaryocytic markers. Importantly, in six CML patient samples studied, silencing BCR-ABL in the lineage depleted enriched stem cell population leads to a decrease in colony-forming capacity. Thus, long-term silencing of BCR-ABL might prove to be a promising alternative approach in CML patients especially for those who do not respond to any other drug treatment.

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Acknowledgements

We thank the FACS facility at CR-UK, London for their excellent support. We also thank Dr Anne Galy and Daniel Compagno for providing the self-inactivating lentiviral and pH1 shuttle vectors. JR would like to thank Rajani Kanth Vangala and family for their continued support and encouragement.

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  1. Haematopoietic Stem Cell Laboratory, Cancer Research UK, London, UK

    J Rangatia & D Bonnet

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  1. J Rangatia
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Correspondence to D Bonnet.

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This work was supported by funding from Cancer Research UK

Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu).

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Rangatia, J., Bonnet, D. Transient or long-term silencing of BCR-ABL alone induces cell cycle and proliferation arrest, apoptosis and differentiation. Leukemia 20, 68–76 (2006). https://doi.org/10.1038/sj.leu.2403999

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