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Kolben, Thomas; Jeschke, Udo; Reimer, Toralf; Karsten, Nora; Schmoeckel, Elisa; Semmlinger, Anna; Mahner, Sven; Harbeck, Nadia and Kolben, Theresa M. (2018): Induction of apoptosis in breast cancer cells in vitro by Fas ligand reverse signaling. In: Journal of Cancer Research and Clinical Oncology, Vol. 144, No. 2: pp. 249-256

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Purpose The Fas-antigen is a cell surface receptor that transduces apoptotic signals into cells. The purpose of this study was to evaluate FasL expression in breast cancer and to elucidate the role of its signaling in different breast cancer cell lines. Methods T47D and MCF7 cells were used and cultured in Dulbecco's modified Eagle's medium. FasL translocation to the membrane was achieved by culturing the cells in the presence of human interferon-gamma (IFN gamma). Translocation was detected by immunofluorescence. The ability of a Fas:Fc fusion protein to trigger apoptosis in these cells was investigated by cell death detection ELISA. After incubation with IFN gamma for 4 h and 18 h, apoptosis was assessed in response to treatment with Fas:Fc. Results Immunofluorescence revealed that the used cell lines were positive for FasL which was increased and changed to more membrane-bound FasL expression after IFN gamma stimulation. After stimulation with 50 IU/ml IFN gamma, Fas:Fc significantly increased MCF7 apoptosis (1.39 +/- 0.06-fold, p = 0.0004) after 18 h. After stimulation with 100 IU/ml, Fas:Fc significantly increased apoptosis both after 4 h (1.49 +/- 0.15-fold, p = 0.018) and 18 h (1.30 +/- 0.06-fold, p = 0.013). In T47D cells this effect was seen after 4 h of stimulation with 50 IU/ml and addition of Fas:Fc (1.6 +/- 0.08-fold, p = 0.03). Conclusion Membrane-bound FasL expression could be induced by IFN gamma in a breast cancer cell model. More importantly, in the presence of IFN gamma the Fas:Fc fusion protein was able to transmit pro-apoptotic signals to T47D and MCF7 cells, significantly inducing apoptosis. The current findings support further in vivo studies regarding FasL activation as a potential target for therapeutic intervention in breast cancer.

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