Kolben Thomas, Jeschke Udo, Reimer Toralf, Karsten Nora, Schmoeckel Elisa, Semmlinger Anna, Mahner Sven, Harbeck Nadia, Kolben Theresa M
Department of Obstetrics and Gynecology, University Hospital, Ludwig-Maximilians-University Munich, Marchioninistr. 15, 81377, Munich, Germany.
Department of Obstetrics and Gynecology, University of Rostock, Suedring 81, 18059, Rostock, Germany.
J Cancer Res Clin Oncol. 2018 Feb;144(2):249-256. doi: 10.1007/s00432-017-2551-y. Epub 2017 Nov 28.
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.
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-γ (IFNγ). 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 IFNγ for 4 h and 18 h, apoptosis was assessed in response to treatment with Fas:Fc.
Immunofluorescence revealed that the used cell lines were positive for FasL which was increased and changed to more membrane-bound FasL expression after IFNγ stimulation. After stimulation with 50 IU/ml IFNγ, 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).
Membrane-bound FasL expression could be induced by IFNγ in a breast cancer cell model. More importantly, in the presence of IFNγ 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.
Fas抗原是一种细胞表面受体，可将凋亡信号传导至细胞内。本研究的目的是评估FasL在乳腺癌中的表达，并阐明其信号传导在不同乳腺癌细胞系中的作用。
使用T47D和MCF7细胞，并在杜氏改良 Eagle培养基中培养。通过在人干扰素-γ（IFNγ）存在的情况下培养细胞，使FasL易位至细胞膜。通过免疫荧光检测易位情况。通过细胞死亡检测ELISA研究Fas:Fc融合蛋白触发这些细胞凋亡的能力。在用IFNγ孵育4小时和18小时后，评估对Fas:Fc处理的凋亡反应。
免疫荧光显示，所用细胞系FasL呈阳性，在IFNγ刺激后FasL增加并转变为更多膜结合形式的表达。在用50 IU/ml IFNγ刺激后，18小时后Fas:Fc显著增加MCF7细胞凋亡（1.39±0.06倍，p = 0.0004）。在用100 IU/ml刺激后，4小时（1.49±0.15倍，p = 0.018）和18小时（1.30±0.06倍，p = 0.013）后Fas:Fc均显著增加细胞凋亡。在T47D细胞中，在用50 IU/ml刺激4小时并添加Fas:Fc后可见此效应（1.6±0.08倍，p = 0.03）。
在乳腺癌细胞模型中，IFNγ可诱导膜结合FasL的表达。更重要的是，在IFNγ存在的情况下，Fas:Fc融合蛋白能够将促凋亡信号传递至T47D和MCF7细胞，显著诱导细胞凋亡。目前的研究结果支持进一步开展关于FasL激活作为乳腺癌治疗干预潜在靶点的体内研究。
