Abstract
MicroRNAs (miRNAs or miRs) regulate diverse normal and abnormal cell functions. We have identified a regulatory pathway in normal megakaryopoiesis, involving the PLZF transcription factor, miR-146a and the SDF-1 receptor CXCR4. In leukaemic cell lines PLZF overexpression downmodulated miR-146a and upregulated CXCR4 protein, whereas PLZF knockdown induced the opposite effects. In vitro assays showed that PLZF interacts with and inhibits the miR-146a promoter, and that miR-146a targets CXCR4 mRNA, impeding its translation. In megakaryopoietic cultures of CD34+ progenitors, PLZF was upregulated, whereas miR-146a expression decreased and CXCR4 protein increased. MiR-146a overexpression and PLZF or CXCR4 silencing impaired megakaryocytic (Mk) proliferation, differentiation and maturation, as well as Mk colony formation. Mir-146a knockdown induced the opposite effects. Rescue experiments indicated that the effects of PLZF and miR-146a are mediated by miR-146a and CXCR4, respectively. Our data indicate that megakaryopoiesis is controlled by a cascade pathway, in which PLZF suppresses miR-146a transcription and thereby activates CXCR4 translation.
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Acknowledgements
We are grateful to M. Fontana, V. Michetti and M. Blasi for editorial assistance. We thank G. Loreto for graphics and A. M. Cerio for technical support. This work was supported by Italy National Grants (Progetto Oncotecnologico, Programma Nazionale Italia-USA in Oncologia) to C.P., institutional grants from the Italian Ministry of Health (Alleanza Contro il Cancro, Programma Straordinario di Ricerca Oncologica 2006) to U.T. and C.L. and Associazione Italiana per la Ricerca sul Cancro (AIRC) to R.F. and F.G.
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C.L., I.S. M.T.Q., E.P., L.P., E.P., M.B., E.R.N., R.F. and U.T. performed experiments; E.B. provided cord-blood samples; F.G. revised the paper. C.L. and C.P. planned and designed the experiments and wrote the paper.
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Labbaye, C., Spinello, I., Quaranta, M. et al. A three-step pathway comprising PLZF/miR-146a/CXCR4 controls megakaryopoiesis. Nat Cell Biol 10, 788–801 (2008). https://doi.org/10.1038/ncb1741
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DOI: https://doi.org/10.1038/ncb1741
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