Knol-Blankevoort Vicky T, Mezzanotte Laura, Rabelink Martijn J W E, Löwik Clemens W G M, Kaijzel Eric L
Department of Radiology, Leiden University Medical Center, Albinusdreef 2, PO Box 9600, 2300RC, Leiden, The Netherlands.
Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.
Methods Mol Biol. 2016;1461:255-70. doi: 10.1007/978-1-4939-3813-1_21.
Here we describe a novel multicolor bioluminescent imaging platform that enables us to simultaneously investigate transcription factor nuclear factor-κB (NF-κB) signalling and apoptosis. We genetically modified the human breast cancer cell line MDA-MB-231 to express green, red, and blue light-emitting luciferases to monitor cell number and viability, NF-κB promoter activity, and to enable specific cell sorting and detection, respectively. Z-DEVD-animoluciferin, the pro-luciferin substrate, was used to determine apoptotic caspase 3/7 activity. We used this multicolored cell line for the in vitro evaluation of natural compounds and in vivo optical imaging of tumor necrosis factor (TNFα)-induced NF-κB activation (Mezzanotte et al., PLoS One 9:e85550, 2014).
在此，我们描述了一种新型的多色生物发光成像平台，该平台使我们能够同时研究转录因子核因子-κB（NF-κB）信号传导和细胞凋亡。我们对人乳腺癌细胞系MDA-MB-231进行基因改造，使其表达绿色、红色和蓝色发光荧光素酶，分别用于监测细胞数量和活力、NF-κB启动子活性，以及进行特定的细胞分选和检测。前荧光素底物Z-DEVD-氨基荧光素用于测定凋亡的半胱天冬酶3/7活性。我们使用这种多色细胞系对天然化合物进行体外评估，并对肿瘤坏死因子（TNFα）诱导的NF-κB激活进行体内光学成像（Mezzanotte等人，《公共科学图书馆·综合》9：e85550，2014年）。
