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In Vitro Functional Study of miR-126 in Leukemia

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MicroRNA and Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 676))

Abstract

MicroRNAs (miRNAs, miRs) are postulated to be important regulators in various cancers, including leukemia. In a large-scale miRNA expression profiling analysis of 435 human miRNAs in 52 acute myeloid leukemia (AML) samples, we found that miR-126 and its minor counterpart in biogenesis, namely, miR-126*, were specifically aberrantly overexpressed in core binding factor (CBF) AMLs including both t(8;21)/AML1-ETO and inv(16)/CBFB-MYH11 samples. Our in vitro gain- and loss-of-function experiments showed that forced expression of miR-126 inhibited apoptosis and increased the viability of AML cells, whereas the opposite effect was observed when endogenous expression of miR-126 was knocked down. In addition, through in vitro colony-forming/replating assays, we demonstrated that forced expression of miR-126 enhanced proliferation and colony-forming/replating capacity of mouse normal bone marrow progenitor cells alone and particularly, in cooperation with AML1-ETO, a fusion gene resulting from t(8;21). Thus, our data shows that miR-126 may play a critical role in the development of CBF leukemias. In the present chapter, the materials and protocols for the study of miR-126 in leukemia are described.

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Acknowledgements

We thank Dr. P. Paul Liu at National Human Genome Research Institute for providing the ME-1 cell line and Dr. Dong-Er Zhang for providing the MIGR1-AE construct. This work was supported in part by National Institutes of Health (NIH) CA127277 (J.C.) and the G. Harold and Leila Y. Mathers Charitable Foundation (J.C.).

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Authors and Affiliations

  1. Section of Hematology and Oncology, Department of Medicine, The University of Chicago, Chicago, IL, USA

    Zejuan Li

  2. Department of Medicine, The University of Chicago, Chicago, IL, USA

    Jianjun Chen

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  1. Zejuan Li
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  2. Jianjun Chen
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  1. , Department of Biological Science, University of Calgary, University Drive N.W. 2500, Calgary, T2N 1N4, Alberta, Canada

    Wei Wu

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Li, Z., Chen, J. (2011). In Vitro Functional Study of miR-126 in Leukemia. In: Wu, W. (eds) MicroRNA and Cancer. Methods in Molecular Biology, vol 676. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-863-8_13

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  • DOI: https://doi.org/10.1007/978-1-60761-863-8_13

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-862-1

  • Online ISBN: 978-1-60761-863-8

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