Redox cycling of 9,10-phenanthrenequinone activates epidermal growth factor receptor signaling through S-oxidation of protein tyrosine phosphatase 1B

J Toxicol Sci. 2020;45(6):349-363. doi: 10.2131/jts.45.349.

Abstract

9,10-Phenanthrenequinone (9,10-PQ) is a polycyclic aromatic hydrocarbon quinone contaminated in diesel exhaust particles and particulate matter 2.5. It is an efficient electron acceptor that induces redox cycling with electron donors, resulting in excessive reactive oxygen species and oxidized protein production in cells. The current study examined whether 9,10-PQ could activate epidermal growth factor receptor (EGFR) signaling in A431 cells through S-oxidation of its negative regulators such as protein tyrosine phosphatase (PTP) 1B. 9,10-PQ oxidized recombinant human PTP1B at Cys215 and inhibited its catalytic activity, an effect that was blocked by catalase (CAT), whereas cis-9,10-dihydroxy-9,10-dihydrophenanthrene (DDP), which lacks redox cycling activity, had no effect on PTP1B activity. Exposure of A431 cells to 9,10-PQ, but not DDP, activated signaling through EGFR and its downstream extracellular signal-regulated kinase 1/2 (ERK1/2), coupled with a decrease of cellular PTP activity. Immunoprecipitation and UPLC-MSE revealed that PTP1B easily undergoes oxidation during exposure of A431 cells to 9,10-PQ. Pretreatment with polyethylene glycol conjugated with CAT (PEG-CAT) abolished 9,10-PQ-generated H2O2 production and significantly blocked the activation of EGFR-ERK1/2 signaling by 9,10-PQ, indicating the involvement of H2O2 in the activation because scavenging agents for hydroxyl radicals had no effect on the redox signal activation. These results suggest that such an air pollutant producing H2O2, activates EGFR-ERK1/2 signaling, presumably through the S-oxidation of PTPs such as PTP1B, and activation of the signal cascade may contribute, at least in part, to cellular responses in A431 cells.

Keywords: 9,10-Phenanthrenequinone; Epidermal growth factor receptor; Hydrogen peroxide; Protein tyrosine phosphatases; Reactive oxygen species; Redox signaling.

MeSH terms

  • Cells, Cultured
  • ErbB Receptors / metabolism*
  • Humans
  • MAP Kinase Signaling System / drug effects
  • Oxidation-Reduction
  • Phenanthrenes / adverse effects*
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1 / metabolism*
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / drug effects*

Substances

  • Phenanthrenes
  • Reactive Oxygen Species
  • 9,10-phenanthrenequinone
  • ErbB Receptors
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1