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Chronic Myeloproliferative Neoplasias

Gfi-1 inhibits proliferation and colony formation of p210BCR/ABL-expressing cells via transcriptional repression of STAT 5 and Mcl-1

Leukemia volume 26, pages 1555–1563 (2012)Cite this article

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Abstract

Expression of the transcription repressor Gfi-1 is required for the maintenance of murine hematopoietic stem cells. In human cells, ectopic expression of Gfi-1 inhibits and RNA interference-mediated Gfi-1 downregulation enhances proliferation and colony formation of p210BCR/ABL expressing cells. To investigate the molecular mechanisms that may explain the effects of perturbing Gfi-1 expression in human cells, Gfi-1-regulated genes were identified by microarray analysis in K562 cells expressing the tamoxifen-regulated Gfi-1-ER protein. STAT 5B and Mcl-1, two genes important for the proliferation and survival of hematopoietic stem cells, were identified as direct and functionally relevant Gfi-1 targets in p210BCR/ABL-transformed cells because: (i) their expression and promoter activity was repressed by Gfi-1 and (ii) when constitutively expressed blocked the proliferation and colony formation inhibitory effects of Gfi-1. Consistent with these findings, genetic or pharmacological inhibition of STAT 5 and/or Mcl-1 markedly suppressed proliferation and colony formation of K562 and CD34+ chronic myelogenous leukemia (CML) cells. Together, these studies suggest that the Gfi-1STAT 5B/Mcl-1 regulatory pathway identified here can be modulated to suppress the proliferation and survival of p210BCR/ABL-transformed cells including CD34+ CML cells.

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Acknowledgements

We thank Dr Michael Andreeff (MD Anderson Hospital, Houston, TX, USA) for the kind gift of the Mcl-1 shRNA/GFP lentiviral plasmid. This work was supported by National Cancer Institute Grants CA 95111 and P01 78890 (BC), by the Cancer Research UK Program Grant C11074/A11008 (TLH) and by the Glasgow Experimental Cancer Medicine Centre (ECMC). MRL was supported by a fellowship of the American-Italian Cancer Foundation (AICF). ARS and GF-A were supported by a fellowship of the Associazione Italiana Ricerca sul Cancro (AIRC). SC was supported, in part, by a fellowship of Fondazione Cassa di Risparmio di Vignola (MO). SAM was supported by a fellowship of the AIRC/Marie Curie Foundation.

Authors contribution

ARS performed most of the molecular studies in K562 cells. SAM performed colony assays and expression studies on primary CML cells. AA and MRL established K562 derivative cell lines and performed colony assays. SA performed microarray analyses. GF-A helped in performing luciferase assays. SC, GM and VF helped in ChIP and real-time PCR analyses. GP provided the −661 +255 Mcl-1-Luc plasmid. LP and TLH provided primary CML samples. BC designed experiments and wrote the paper.

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Authors and Affiliations

  1. Department of Cancer Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA

    A R Soliera, S A Mariani, A Audia, M R Lidonnici, S Addya & B Calabretta

  2. Department of Biomedical Sciences, University of Modena, Modena, Italy

    A R Soliera, G Ferrari-Amorotti, S Cattelani, G Manzotti, V Fragliasso & B Calabretta

  3. Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan Comprehensive Cancer Center, Ann Arbor, MI, USA

    L Peterson

  4. Department of Biology, University of Bologna, Bologna, Italy

    G Perini

  5. Paul O’Gorman Leukaemia Research Centre, University of Glasgow and Gartnavel General Hospital, Glasgow, UK

    T L Holyoake

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Correspondence to B Calabretta.

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Soliera, A., Mariani, S., Audia, A. et al. Gfi-1 inhibits proliferation and colony formation of p210BCR/ABL-expressing cells via transcriptional repression of STAT 5 and Mcl-1. Leukemia 26, 1555–1563 (2012). https://doi.org/10.1038/leu.2012.19

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