PMID- 25886616 OWN - NLM STAT- MEDLINE DCOM- 20160119 LR - 20210109 IS - 1756-8722 (Electronic) IS - 1756-8722 (Linking) VI - 8 DP - 2015 Mar 21 TI - EVI1 promotes tumor growth via transcriptional repression of MS4A3. PG - 28 LID - 10.1186/s13045-015-0124-6 [doi] LID - 28 AB - BACKGROUND: The transcription factor Ecotropic Virus Integration site 1 (EVI1) regulates cellular proliferation, differentiation, and apoptosis, and its overexpression contributes to an aggressive course of disease in myeloid leukemias and other malignancies. Notwithstanding, knowledge about the target genes mediating its biological and pathological functions remains limited. We therefore aimed to identify and characterize novel EVI1 target genes in human myeloid cells. METHODS: U937T_EVI1, a human myeloid cell line expressing EVI1 in a tetracycline regulable manner, was subjected to gene expression profiling. qRT-PCR was used to confirm the regulation of membrane-spanning-4-domains subfamily-A member-3 (MS4A3) by EVI1. Reporter constructs containing various parts of the MS4A3 upstream region were employed in luciferase assays, and binding of EVI1 to the MS4A3 promoter was investigated by chromatin immunoprecipitation. U937 derivative cell lines experimentally expressing EVI1 and/or MS4A3 were generated by retroviral transduction, and tested for their tumorigenicity by subcutaneous injection into severe combined immunodeficient mice. RESULTS: Gene expression microarray analysis identified 27 unique genes that were up-regulated, and 29 unique genes that were down-regulated, in response to EVI1 induction in the human myeloid cell line U937T. The most strongly repressed gene was MS4A3, and its down-regulation by EVI1 was confirmed by qRT-PCR in additional, independent experimental model systems. MS4A3 mRNA levels were also negatively correlated with those of EVI1 in several published AML data sets. Reporter gene assays and chromatin immunoprecipitation showed that EVI1 regulated MS4A3 via direct binding to a promoter proximal region. Experimental re-expression of MS4A3 in an EVI1 overexpressing cell line counteracted the tumor promoting effect of EVI1 in a murine xenograft model by increasing the rate of apoptosis. CONCLUSIONS: Our data reveal MS4A3 as a novel direct target of EVI1 in human myeloid cells, and show that its repression plays a role in EVI1 mediated tumor aggressiveness. FAU - Heller, Gerwin AU - Heller G AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. gerwin.heller@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. gerwin.heller@meduniwien.ac.at. FAU - Rommer, Anna AU - Rommer A AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. apm.rommer@gmail.com. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. apm.rommer@gmail.com. FAU - Steinleitner, Katarina AU - Steinleitner K AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. katarina.steinleitner@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. katarina.steinleitner@meduniwien.ac.at. FAU - Etzler, Julia AU - Etzler J AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. julia.etzler@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. julia.etzler@meduniwien.ac.at. FAU - Hackl, Hubert AU - Hackl H AD - Biocenter, Division of Bioinformatics, Medical University of Innsbruck, Innrain 80, 6020, Innsbruck, Austria. hubert.hackl@i-med.ac.at. FAU - Heffeter, Petra AU - Heffeter P AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. petra.heffeter@meduniwien.ac.at. AD - Department of Medicine I, Institute of Cancer Research, and Research Platform "Translational Cancer Therapy Research", Borschkegasse 8A, 1090, Vienna, Austria. petra.heffeter@meduniwien.ac.at. FAU - Tomasich, Erwin AU - Tomasich E AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. erwin.tomasich@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. erwin.tomasich@meduniwien.ac.at. FAU - Filipits, Martin AU - Filipits M AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. martin.filipits@meduniwien.ac.at. AD - Department of Medicine I, Institute of Cancer Research, and Research Platform "Translational Cancer Therapy Research", Borschkegasse 8A, 1090, Vienna, Austria. martin.filipits@meduniwien.ac.at. FAU - Steinmetz, Birgit AU - Steinmetz B AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. birgit.steinmetz@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. birgit.steinmetz@meduniwien.ac.at. FAU - Topakian, Thais AU - Topakian T AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. thais.topakian@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. thais.topakian@meduniwien.ac.at. FAU - Klingenbrunner, Simone AU - Klingenbrunner S AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. sim.kling@gmx.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. sim.kling@gmx.at. FAU - Ziegler, Barbara AU - Ziegler B AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. barbara.ziegler@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. barbara.ziegler@meduniwien.ac.at. FAU - Spittler, Andreas AU - Spittler A AD - Core Facility Flow Cytometry & Surgical Research Laboratories, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. andreas.spittler@meduniwien.ac.at. FAU - Zochbauer-Muller, Sabine AU - Zochbauer-Muller S AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. sabine.zoechbauer-mueller@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. sabine.zoechbauer-mueller@meduniwien.ac.at. FAU - Berger, Walter AU - Berger W AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. walter.berger@meduniwien.ac.at. AD - Department of Medicine I, Institute of Cancer Research, and Research Platform "Translational Cancer Therapy Research", Borschkegasse 8A, 1090, Vienna, Austria. walter.berger@meduniwien.ac.at. FAU - Wieser, Rotraud AU - Wieser R AD - Department of Medicine I, Medical University of Vienna, Wahringer Gurtel 18-20, 1090, Vienna, Austria. rotraud.wieser@meduniwien.ac.at. AD - Comprehensive Cancer Center of the Medical University of Vienna, Vienna, Austria. rotraud.wieser@meduniwien.ac.at. LA - eng GR - P 24130/FWF_/Austrian Science Fund FWF/Austria PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150321 PL - England TA - J Hematol Oncol JT - Journal of hematology & oncology JID - 101468937 RN - 0 (Cell Cycle Proteins) RN - 0 (DNA-Binding Proteins) RN - 0 (MDS1 and EVI1 Complex Locus Protein) RN - 0 (MECOM protein, human) RN - 0 (MS4A3 protein, human) RN - 0 (Membrane Proteins) RN - 0 (Transcription Factors) SB - IM MH - Animals MH - Cell Cycle Proteins/genetics/*metabolism MH - Cell Line, Tumor MH - *Cell Proliferation/physiology MH - Chromatin Immunoprecipitation MH - DNA-Binding Proteins/genetics/*metabolism MH - Fluorescent Antibody Technique MH - Gene Expression Regulation, Neoplastic/*physiology MH - Gene Knockdown Techniques MH - Heterografts MH - Humans MH - Immunohistochemistry MH - Leukemia, Myeloid/genetics/metabolism/*pathology MH - MDS1 and EVI1 Complex Locus Protein MH - Male MH - Membrane Proteins/genetics/*metabolism MH - Mice MH - Mice, SCID MH - Oligonucleotide Array Sequence Analysis MH - Proto-Oncogenes/genetics MH - Reverse Transcriptase Polymerase Chain Reaction MH - Transcription Factors/genetics/*metabolism MH - *Transcription, Genetic PMC - PMC4389965 EDAT- 2015/04/18 06:00 MHDA- 2016/01/20 06:00 PMCR- 2015/03/21 CRDT- 2015/04/18 06:00 PHST- 2014/08/12 00:00 [received] PHST- 2015/02/26 00:00 [accepted] PHST- 2015/04/18 06:00 [entrez] PHST- 2015/04/18 06:00 [pubmed] PHST- 2016/01/20 06:00 [medline] PHST- 2015/03/21 00:00 [pmc-release] AID - 10.1186/s13045-015-0124-6 [pii] AID - 124 [pii] AID - 10.1186/s13045-015-0124-6 [doi] PST - epublish SO - J Hematol Oncol. 2015 Mar 21;8:28. doi: 10.1186/s13045-015-0124-6.