Distinct responses of compartmentalized glutathione redox potentials to pharmacologic quinones targeting NQO1

Biochem Biophys Res Commun. 2017 Jan 29;483(1):680-686. doi: 10.1016/j.bbrc.2016.12.082. Epub 2016 Dec 14.

Abstract

Deoxynyboquinone (DNQ), a potent novel quinone-based antineoplastic agent, selectively kills solid cancers with overexpressed cytosolic NAD(P)H:quinone oxidoreductase-1 (NQO1) via excessive ROS production. A genetically encoded redox-sensitive probe was used to monitor intraorganellar glutathione redox potentials (EGSH) as a direct indicator of cellular oxidative stress following chemotherapeutic administration. Beta-lapachone (β-lap) and DNQ-induced spatiotemporal redox responses were monitored in human lung A549 and pancreatic MIA-PaCa-2 adenocarcinoma cells incubated with or without dicumarol and ES936, potent NQO1 inhibitors. Immediate oxidation of EGSH in both the cytosol and mitochondrial matrix was observed in response to DNQ and β-lap. The DNQ-induced cytosolic oxidation was fully prevented with NQO1 inhibition, whereas mitochondrial oxidation in A549 was NQO1-independent in contrast to MIA-PaCa-2 cells. However, at pharmacologic concentrations of β-lap both quinone-based substrates directly oxidized the redox probe, a possible sign of off-target reactivity with cellular thiols. Together, these data provide new evidence that DNQ's direct and discerning NQO1 substrate specificity underlies its pharmacologic potency, while β-lap elicits off-target responses at its effective doses.

Keywords: DNQ and β-lapachone; Lung and pancreatic cancer; NQO1; Redox homeostasis; roGFP2 redox probe.

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Biosensing Techniques
  • Cell Line, Tumor
  • Cytosol / drug effects
  • Cytosol / metabolism
  • Dicumarol / pharmacology
  • Fluorescent Dyes / analysis
  • Glutaredoxins / analysis
  • Glutaredoxins / genetics
  • Glutathione / analysis
  • Glutathione / metabolism*
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / genetics
  • Humans
  • Indolequinones / pharmacology
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • Molecular Imaging
  • Molecular Probes / genetics
  • Molecular Targeted Therapy
  • NAD(P)H Dehydrogenase (Quinone) / antagonists & inhibitors
  • NAD(P)H Dehydrogenase (Quinone) / metabolism*
  • Naphthoquinones / metabolism
  • Oxidation-Reduction / drug effects
  • Oxidative Stress / drug effects*
  • Quinones / pharmacology*
  • Reactive Oxygen Species / metabolism
  • Substrate Specificity

Substances

  • 5-methoxy-1,2-dimethyl-3-((4-nitrophenoxy)methyl)indole-4,7-dione
  • Antineoplastic Agents
  • Fluorescent Dyes
  • Glutaredoxins
  • Indolequinones
  • Molecular Probes
  • Naphthoquinones
  • Quinones
  • Reactive Oxygen Species
  • deoxynyboquinone
  • Green Fluorescent Proteins
  • beta-lapachone
  • Dicumarol
  • NAD(P)H Dehydrogenase (Quinone)
  • NQO1 protein, human
  • Glutathione