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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands.
Laura
L
Mezzanotte
Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands.
Na
N
An
Department of Endocrinology, Leiden University Medical Center, Leiden, The Netherlands.
Isabel M
IM
Mol
Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands.
Clemens W G M
CW
Löwik
Department of Radiology, Leiden University Medical Center, Leiden, The Netherlands.
Eric L
EL
Kaijzel
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045744:Cell Line, Tumor; D018890:Chemoprevention; D005260:Female; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D021621:Imaging, Three-Dimensional; D016086:Lentivirus / Q000235:genetics; D049449:Luminescence; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D053978:Pentacyclic Triterpenes; D011401:Promoter Regions, Genetic / Q000235:genetics; D013997:Time Factors; D014315:Triterpenes / Q000494:pharmacology / Q000627:therapeutic use; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D016328:NF-kappa B; D053978:Pentacyclic Triterpenes; D014315:Triterpenes; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3; D053179:Caspase 7; C050414:celastrol