Benzoquinone activates the ERK/MAPK signaling pathway via ROS production in HL-60 cells

Toxicology. 2005 May 5;209(3):279-87. doi: 10.1016/j.tox.2004.12.035.

Abstract

Benzene (BZ) is a class I carcinogen and its oxidation to reactive intermediates is a prerequisite of hematoxicity and myelotoxicity. The generated metabolites include hydroquinone, which is further oxidized to the highly reactive 1,4-benzoquinone (BQ) in bone marrow. Therefore, we explored the mechanisms underlying BQ-induced HL-60 cell proliferation by studying the role of BQ-induced reactive oxygen species (ROS) in the activation of the ERK-MAPK signaling pathway. BQ treatment (0.01-30 microM) showed that doses below 10 microM did not significantly reduce viability. ROS production after 3 microM BQ treatment increased threefold; however, catalase addition reduced ROS generation to basal levels. FACS analysis showed that BQ induced a fivefold increase in the proportion of cells in S-phase. We also observed a high proportion of Bromodeoxyuridine (BrdU) stained cells, indicating a higher DNA synthesis rate. BQ also produced rapid and prolonged phosphorylation of ERK1/2 proteins. Simultaneous treatment with catalase or PD98059, a potent MEK protein inhibitor, reduced cell recruitment into the S-phase and also abolished the ERK1/2 protein phosphorylation induced by BQ, suggesting that MEK/ERK is an important pathway involved in BQ-induced ROS mediated proliferation. The prolonged activation of ERK1/2 contributes to explain the increased S-phase cell recruitment and to understand the leukemogenic processes associated with exposure to benzene metabolites. Thus, the possible mechanism by which BQ induce HL-60 cells to enter the cell cycle and proliferate is linked to ROS production and its growth promoting effects by specific activation of regulating genes known to be activated by redox mechanisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzoquinones / toxicity*
  • Bromodeoxyuridine
  • Butadienes / pharmacology
  • Catalase / pharmacology
  • Cell Proliferation / drug effects*
  • Cell Survival / drug effects
  • Enzyme Inhibitors / pharmacology
  • Flavonoids / pharmacology
  • HL-60 Cells
  • Humans
  • MAP Kinase Kinase Kinase 1 / antagonists & inhibitors
  • MAP Kinase Kinase Kinase 1 / metabolism
  • MAP Kinase Kinase Kinase 2 / antagonists & inhibitors
  • MAP Kinase Kinase Kinase 2 / metabolism
  • Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
  • Mitogen-Activated Protein Kinase 1 / metabolism*
  • Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
  • Mitogen-Activated Protein Kinase 3 / metabolism*
  • Nitriles / pharmacology
  • Phosphorylation
  • Reactive Oxygen Species / metabolism*
  • Signal Transduction

Substances

  • Benzoquinones
  • Butadienes
  • Enzyme Inhibitors
  • Flavonoids
  • Nitriles
  • Reactive Oxygen Species
  • U 0126
  • quinone
  • Catalase
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • MAP Kinase Kinase Kinase 1
  • MAP Kinase Kinase Kinase 2
  • Bromodeoxyuridine
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one