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Gene profiling of growth factor independence 1B gene (Gfi-1B) in leukemic cells

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Abstract

To investigate the molecular effects of growth factor independence 1B (Gfi-1B), a transcription factor essential for the development of hematopoietic cells and differentiation of erythroid and megakaryocytic lineages, the naturally Gfi-1B overexpressing cell line K562 was cultured in the presence of Gfi-1B target-specific small interfering RNA (siRNA). SiRNA treatment significantly knocked down Gfi-1B expression with an efficiency of nearly 90%. Analysis of the siRNA silencing protocol by colony-forming units ensured that it was not cytotoxic. Samples from Gfi-1B overexpressing cells and cells with knocked-down Gfi-1B were analyzed by oligonucleotide microarray technology and based upon rigorous statistical analysis of the data; relevant genes were chosen for confirmation by reserve transcriptase-polymerase chain reaction, including MYC/MYCBP and CDKN1A. Interestingly, transcripts within components of the signalling cascade of immune cells (PLD1, LAMP1, HSP90, IL6ST), of the tyrosine kinase pathway (TPR, RAC3) and of the transcription factors (RAC3, CEP290, JEM-1, ATR, MYC, SMC3, RARA, RBBP6) were found to be differentially expressed in Gfi-1B overexpressing cells compared to controls. Individual genes such as ZDHHC17, DMXL1, ZNF292 were found to be upregulated in Gfi-1B overexpressing cells. In addition, down-regulated transcripts showed cell signaling transcripts for several chemokine gene members including GNAL, CXCL5, GNL3L, GPR65, TMEM30, BCL11B and transcription factors (GTF2H3, ATXN3). In conclusion, several essential cell signalling factors, as well as transcriptional and post-translational regulation genes were differentially expressed in cells that overexpressed Gfi-1B compared to control cells with knocked-down Gfi-1B. Our data indicate that Gfi-1B signalling is important for commitment and maturation of hematopoietic cell populations.

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Acknowledgments

The authors would like to thank Liana Baatsch, Melanie Kroll, Silke Gottwald, Katja Schneider, Christiane Schary, and Ines Riepenhoff for their excellent technical performance of the MA analyses, PCR analyses, CFU assays and siRNA experiments.

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Authors and Affiliations

  1. Department of Bone Marrow Transplantation, University Hospital of Duisburg-Essen, Hufelandstr. 55, 45122, Essen, Germany

    Michael Koldehoff, Dietrich W. Beelen & Ahmet H. Elmaagacli

  2. Immunology of Allogeneic Bone Marrow Transplantation, Memorial Sloan Kettering, New York City, USA

    Johannes L. Zakrzewski

  3. Institute of Cell Biology, University of Duisburg-Essen, Essen, Germany

    Ludger Klein-Hitpass

Authors
  1. Michael Koldehoff
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  2. Johannes L. Zakrzewski
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  3. Ludger Klein-Hitpass
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  4. Dietrich W. Beelen
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  5. Ahmet H. Elmaagacli
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Corresponding author

Correspondence to Michael Koldehoff.

Additional information

This work was supported in part by grants of the Förderverein Essener Tumorklinik e.V. and the Kulturstiftung Essen e.V.

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Koldehoff, M., Zakrzewski, J.L., Klein-Hitpass, L. et al. Gene profiling of growth factor independence 1B gene (Gfi-1B) in leukemic cells. Int J Hematol 87, 39–47 (2008). https://doi.org/10.1007/s12185-007-0013-z

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  • DOI: https://doi.org/10.1007/s12185-007-0013-z

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