caa a22 4500 60617494X CHVBK 20210128100740.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00018-015-1910-6 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1910-6 Quantification of cell fusion events human breast cancer cells and breast epithelial cells using a Cre-LoxP-based double fluorescence reporter system [Elektronische Daten] [Marieke Mohr, Songül Tosun, Wolfgang Arnold, Frank Edenhofer, Kurt Zänker, Thomas Dittmar] The biological phenomenon of cell fusion plays an important role in several physiological processes, like fertilization, placentation, or wound healing/tissue regeneration, as well as pathophysiological processes, such as cancer. Despite this fact, considerably less is still known about the factors and conditions that will induce the merging of two plasma membranes. Inflammation and proliferation has been suggested as a positive trigger for cell fusion, but it remains unclear, which of the factor(s) of the inflamed microenvironment are being involved. To clarify this we developed a reliable assay to quantify the in vitro fusion frequency of cells using a fluorescence double reporter vector (pFDR) containing a LoxP-flanked HcRed/DsRed expression cassette followed by an EGFP expression cassette. Because cell fusion has been implicated in cancer progression four human breast cancer cell lines were stably transfected with a pFDR vector and were co-cultured with the stably Cre-expressing human breast epithelial cell line. Cell fusion is associated with a Cre-mediated recombination resulting in induction of EGFP expression in hybrid cells, which can be quantified by flow cytometry. By testing a panel of different cytokines, chemokines, growth factors and other compounds, including exosomes, under normoxic and hypoxic conditions our data indicate that the proinflammatory cytokine TNF-α together with hypoxia is a strong inducer of cell fusion in human MDA-MB-435 and MDA-MB-231 breast cancer cells. Springer Basel, 2015 Cell fusion nationallicence Breast cancer nationallicence Cre-LoxP recombination nationallicence Flow cytometry nationallicence Mohr Marieke Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut Tosun Songül Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut Arnold Wolfgang Department of Biological and Material Sciences in Dentistry, School of Dentistry, Faculty of Health, Witten/Herdecke University, Witten, Germany aut Edenhofer Frank Stem Cell and Regenerative Medicine Group, Institute of Anatomy and Cell Biology, Julius-Maximilians-University Würzburg, Würzburg, Germany aut Zänker Kurt Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut Dittmar Thomas Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut Cellular and Molecular Life Sciences Springer Basel 72/19(2015-10-01), 3769-3782 1420-682X 72:19<3769 2015 72 18 https://doi.org/10.1007/s00018-015-1910-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1910-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mohr Marieke Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut NATIONALLICENCE 700 1- Tosun Songül Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut NATIONALLICENCE 700 1- Arnold Wolfgang Department of Biological and Material Sciences in Dentistry, School of Dentistry, Faculty of Health, Witten/Herdecke University, Witten, Germany aut NATIONALLICENCE 700 1- Edenhofer Frank Stem Cell and Regenerative Medicine Group, Institute of Anatomy and Cell Biology, Julius-Maximilians-University Würzburg, Würzburg, Germany aut NATIONALLICENCE 700 1- Zänker Kurt Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut NATIONALLICENCE 700 1- Dittmar Thomas Institute of Immunology and Experimental Oncology, Center for Biomedical Education and Research, Witten/Herdecke University, Stockumer Str. 10, 58448, Witten, Germany aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/19(2015-10-01), 3769-3782 1420-682X 72:19<3769 2015 72 18