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Anal Chem. 2013 Aug 6;85(15):7542-9. doi: 10.1021/ac401569j. Epub 2013 Jul 23.

Reporter protein complementation imaging assay to screen and study Nrf2 activators in cells and living animals.

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1
SRM Research Institute, SRM University, Kattankulathur-603203, Tamilnadu, India.

Abstract

NF-E2-related factor-2 (Nrf2) activators promote cellular defense mechanism and facilitate disease prevention associated with oxidative stress. In the present study, Nrf2 activators were identified using cell-based luciferase enzyme fragment complementation (EFC) assay, and the mechanism of Nrf2 activation was studied by molecular imaging. Among the various Nrf2 activators tested, pterostilbene (PTS) showed effective Nrf2 activation, as seen by luminometric screening, and validation in a high throughput-intact cell-imaging platform. Further, PTS increased the expression of Nrf2 downstream target genes, which was confirmed using luciferase reporter driven by ARE-NQO1 and ARE-GST1 promoters. Daily administration of PTS disturbed Nrf2/Keap1 interaction and reduced complemented luciferase signals in HEK293TNKS mouse tumor xenografts. This study reveals the potentials of Nrf2 activators as chemosensitizing agents' for therapeutic intervention in cancer treatment. Hence, the validated assay can be used to evaluate the identified activators preclinically in small animal models by noninvasive molecular imaging approach.

PMID:
23826874
PMCID:
PMC3759980
DOI:
10.1021/ac401569j
[Indexed for MEDLINE]
Free PMC Article
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