Induction of heme oxygenase-1 by phenylarsine oxide. Studies in cultured primary liver cells

Mol Cell Biochem. 2001 Oct;226(1-2):17-26.

Abstract

Heme oxygenase-1, the major inducible isoform of heme oxygenase (HO), can be induced by heme and numerous other physical and chemical factors, many of which cause cellular 'stress'. This has led to the realization that HO-1 is a major highly conserved stress or heat shock protein. Recent work has implicated activation of mitogen-activated protein kinases and other kinases in the mechanism of induction of HO-1, and suggested that signal transduction pathways through tyrosine kinases are involved in induction of HO-1 gene expression by stress inducers. We hypothesized that phenylarsine oxide (PAO), an inhibitor of protein tyrosine phosphatases (PTPs), might up-regulate the HO-1 gene. Here, we show that a remarkably brief (1-15 min) exposure of normal hepatocytes to low concentrations (0.5-3 microM) of PAO produces a marked increase in mRNA and protein of HO-1. This increase is comparable to the level obtained by addition of heme (20 microM), and occurs without producing changes in cellular glutathione levels or stabilization of HO-1 message. Preincubation of cells with inhibitors of protein synthesis decreased the ability of PAO to increase levels of HO-1 mRNA, suggesting that the inductive effect requires de novo protein synthesis. Addition of thiol donors abrogated the PAO-mediated induction of HO-1 in a dose dependent fashion. Addition of genistein, a tyrosine kinase inhibitor, blunted the induction produced by both PAO and heme. After brief incubations with PAO or heme, cell extracts showed comparable increases in levels of protein tyrosine phosphorylation in general, and specifically in ZAP70 kinase. Our results are consistent with the proposition that induction of HO-1 by PAO involves inhibition of specific PTP(s), and that the mechanisms of induction of HO-1 by PAO and by heme may share some common pathways.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actins / metabolism
  • Animals
  • Arsenicals / metabolism*
  • Blotting, Western
  • Cells, Cultured
  • Chick Embryo
  • Dactinomycin / pharmacology
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / pharmacology
  • Genistein / pharmacology
  • Glutathione / metabolism
  • Heme Oxygenase (Decyclizing) / metabolism*
  • Heme Oxygenase-1
  • Hepatocytes / metabolism
  • L-Lactate Dehydrogenase / metabolism
  • Liver / cytology*
  • Phosphorylation
  • Protein Isoforms
  • Protein-Tyrosine Kinases / antagonists & inhibitors
  • Protein-Tyrosine Kinases / metabolism
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Stress, Physiological
  • Sulfhydryl Compounds / chemistry
  • Time Factors
  • Up-Regulation
  • ZAP-70 Protein-Tyrosine Kinase

Substances

  • Actins
  • Arsenicals
  • Enzyme Inhibitors
  • Protein Isoforms
  • RNA, Messenger
  • Sulfhydryl Compounds
  • oxophenylarsine
  • Dactinomycin
  • Genistein
  • L-Lactate Dehydrogenase
  • Heme Oxygenase (Decyclizing)
  • Heme Oxygenase-1
  • Protein-Tyrosine Kinases
  • ZAP-70 Protein-Tyrosine Kinase
  • Glutathione