An improved cell-permeable fluorogenic substrate as the basis for a highly sensitive test for NAD(P)H quinone oxidoreductase 1 (NQO1) in living cells.
Free Radic Biol Med
; 116: 141-148, 2018 02 20.
Article
en En
| MEDLINE
| ID: mdl-29325897
ABSTRACT
NAD(P)Hquinone oxidoreductase 1 (NQO1) is a flavoenzyme upregulated in response to oxidative stress and in some cancers. Its upregulation by compounds has been used as an indicator of their potential anti-cancer properties. In this study we have designed, produced and tested a fluorogenic coumarin conjugate which selectively releases highly fluorescent 4-methylumbelliferone (4-MU) in the presence of NQO1. It was found that measuring 4-MU release rapidly and specifically quantitated NQO1 levels in vitro and in live cells. Both the substrate and its products freely perfused through cell membranes and were non-toxic. The substrate was very specific with low background, and the assay itself could be done in less than 10minutes. This is the first assay to allow the quantitation of NQO1 in live cells which can then be retained for further experiments.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Biomarcadores
/
Membrana Celular
/
NAD(P)H Deshidrogenasa (Quinona)
/
Cumarinas
/
Colorantes Fluorescentes
/
Neoplasias
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
En
Revista:
Free Radic Biol Med
Asunto de la revista:
BIOQUIMICA
/
MEDICINA
Año:
2018
Tipo del documento:
Article