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MicroRNA expression in lymphocyte development and malignancy

Leukemia volume 22, pages 1440–1446 (2008)Cite this article

MicroRNAs are a recently discovered class of naturally occurring short noncoding RNA molecules that negatively regulate eukaryotic gene expression through binding to complementary sequences in the 3′-UTR of target mRNA. There is rapidly accumulating evidence to suggest that dysfunctional expression of microRNAs is a common feature of hematological malignancy.1 With the exception perhaps of chronic lymphocytic leukemia (CLL) however, the role of microRNAs in hematological malignancies is not well understood; in particular, to date there has been no systematic identification of aberrantly expressed microRNAs in lymphoma. To address this, we used microarray analysis to look at microRNA expression in a wide range of hematological cell lines (n=40) including all major types of B- and T-cell lymphoma (n=29), nonmalignant lymphoproliferative disorder, persistent polyclonal B-cell lymphocytosis (PPBL; n=3), T-cell acute lymphoblastic leukemia (T-ALL; n=6) and acute myeloid leukemia (AML; n=2; Table 1). To compare these samples with their normal counterpart cells of origin, as well as looking at the role of microRNAs in lymphocyte development, we also examined expression in the major B-lymphocyte developmental stages and mature T-cell subsets (n=12; Table 1).

Table 1 Cell lines and lymphocyte subsets used for microRNA expression analysis
Full size table

Samples (approximately 107 cells) were first purified into microRNA-enriched fractions using miREasy columns from Qiagen (Crawley, UK) and 500 ng labeled with either Cy3 or Cy5 using the Array 900microRNA RT kit from Genisphere (Hatfield, PA, USA). Tonsillar material, pooled from 12 healthy individuals, was used as a common reference for experiments using a dye-balanced design. Samples were hybridized to microarrays (μRNA microarray) that include the complete human and EBV-derived sense mature microRNA probe sets (MirBase v.9.2) spotted in quadruplicate nonadjacently. Probe sequences are available at www.microRNAworld.com. The sensitivity, specificity and reproducibility of the arrays were tested as described (Supplementary Figures S1–S3). Levels of selected microRNAs were measured by quantitative RT–PCR (qRT-PCR) as previously described2 using U6 as a control.

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Acknowledgements

This work was funded by grants from the Leukemia Research Fund (CHL, HMD, PJB, JB and JSW), the Julian Starmer-Smith Memorial Fund (CHL and CDOC) and the Medical Research Council (SS). The microarray printing infrastructure was established with financial support from the EPA Cephalosporin Fund. We are very grateful to Helen Roberton, Linden Lyne, Dr Karen Pulford and Dr Alison Banham for cell line material. We declare no potential conflicts of interest.

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Authors and Affiliations

  1. Nuffield Department of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford, UK

    C H Lawrie, S Palazzo, H M Dunlop, C D O Cooper, P J Brown, F Pezzella, J Boultwood & J S Wainscoat

  2. Sir William Dunn School of Pathology, University of Oxford, Oxford, UK

    N J Saunders

  3. Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK

    S Soneji & T Enver

  4. Laboratoire d'Hématologie, CHU de Caen, Caen, France

    X Troussard

  5. Déparement de Genetique, Laboratoire Pasteur, Cerba, France

    H Mossafa

  6. Department of Haematology, John Radcliffe Hospital, Oxford, UK

    C S R Hatton

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  1. C H Lawrie
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Correspondence to C H Lawrie.

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

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Lawrie, C., Saunders, N., Soneji, S. et al. MicroRNA expression in lymphocyte development and malignancy. Leukemia 22, 1440–1446 (2008). https://doi.org/10.1038/sj.leu.2405083

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