Development of a Multicolor Bioluminescence Imaging Platform to Simultaneously Investigate Transcription Factor NF-κB Signaling and Apoptosis

Methods Mol Biol. 2016:1461:255-70. doi: 10.1007/978-1-4939-3813-1_21.

Abstract

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).

Keywords: Apoptosis; Bioluminescence; Breast cancer; Gaussia; Multicolor; NF-κB; TNFα.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism
  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cell Line, Tumor
  • Disease Models, Animal
  • Female
  • Gene Expression
  • Genes, Reporter
  • Genetic Vectors / genetics
  • Humans
  • Luminescent Measurements / methods*
  • Mice
  • Molecular Imaging / methods*
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • Recombinant Fusion Proteins
  • Signal Transduction / drug effects*
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • Xenograft Model Antitumor Assays

Substances

  • NF-kappa B
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • Caspase 3
  • Caspase 7