A new multicolor bioluminescence imaging platform to investigate NF-κB activity and apoptosis in human breast cancer cells

• Published in: PloS one 2014, Vol.9 (1), p.e85550-e85550
• Main Authors: Mezzanotte, Laura, An, Na, Mol, Isabel M, Löwik, Clemens W G M, Kaijzel, Eric L
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Apoptosis; Apoptosis - drug effects; Biology; Bioluminescence; Breast cancer; Breast Neoplasms - drug therapy; Breast Neoplasms - enzymology; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cancer; Cancer therapies; Caspase; Caspase 3 - metabolism; Caspase 7 - metabolism; Caspase-3; Cell Line, Tumor; Cell number; Chemoprevention; Cloning; Curcumin; Disease prevention; Drug development; Drugs; Endocrinology; Evaluation; Female; Gaussia; Gene expression; Genetic modification; Genetic Vectors - metabolism; Humans; Imaging; Imaging, Three-Dimensional; In vivo methods and tests; Laboratory animals; Lentivirus - genetics; Light emitting; Luciferin; Luminescence; Medical research; Medicine; Metastasis; Mice; Nanoparticles; NF-kappa B - metabolism; NF-κB protein; Optimization; Promoter Regions, Genetic - genetics; Proteins; Resveratrol; Screening; Signal transduction; Signaling; Studies; Substrates; Time Factors; Triterpenes - pharmacology; Triterpenes - therapeutic use; Tumor Necrosis Factor-alpha - pharmacology; Tumor necrosis factor-α; Viability
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0085550

Evaluation of novel drugs for clinical development depends on screening technologies and informative preclinical models. Here we developed a multicolor bioluminescent imaging platform to simultaneously investigate transcription factor NF-κB signaling and apoptosis. The human breast cancer cell line (MDA-MB-231) was genetically modified to express green, red and blue light emitting luciferases to monitor cell number and viability, NF-κB promoter activity and to perform specific cell sorting and detection, respectively. The pro-luciferin substrate Z-DEVD-animoluciferin was employed to determine apoptotic caspase 3/7 activity. We used the cell line for the in vitro evaluation of natural compounds and in vivo optical imaging of tumor necrosis factor TNFα-induced NF-κB activation. Celastrol, resveratrol, sulphoraphane and curcumin inhibited the NF-κB promoter activity significantly and in a dose dependent manner. All compounds except resveratrol induced caspase 3/7 dependent apoptosis. Multicolor bioluminescence in vivo imaging allowed the investigation of tumor growth and NF-κB induction in a mouse model of breast cancer. Our new method provides an imaging platform for the identification, validation, screening and optimization of compounds acting on NF-κB signaling and apoptosis both in vitro and in vivo.