Transfection of mGSTA4 in HL-60 cells protects against 4-hydroxynonenal-induced apoptosis by inhibiting JNK-mediated signaling

Arch Biochem Biophys. 2001 Aug 15;392(2):197-207. doi: 10.1006/abbi.2001.2452.


The mammalian alpha-class glutathione S-transferase (GST) isozymes mGSTA4-4, rGSTA4-4, and hGSTA4-4 are known to utilize 4-hydroxynonenal (4HNE) as a preferred substrate. During the present studies, we have examined the effect of transfecting human myeloid HL-60 cells with mGSTA4, on 4-HNE-induced apoptosis and the associated signaling mechanisms. Results of these studies show that treatment of the wild-type or vector-only-transfected HL-60 cells with 20 microM 4-HNE caused apoptosis within 2 h. The cells transfected with mGSTA4 did not undergo apoptosis under these conditions even after 4 h. In the wild-type and vector-transfected cells, apoptosis was preceded by JNK activation and c-Jun phosphorylation within 30 min, and an increase in AP-1 binding within 2 h of treatment with 20 microM 4-HNE. In mGSTA4-transfected cells, JNK activation and c-Jun phosphorylation were observed after 1 h, and increased AP-1 binding was observed after 8 h under these conditions. In the control cells, 20 microM 4-HNE caused caspase 3 activation and poly(ADP-ribose) polymerase cleavage within 2 h, while in mGSTA4-transfected cells, a lesser degree of these effects was observed even after 8 h. Transfection with mGSTA4 also provided protection to the cells from 4-HNE and doxorubicin cytotoxicity (1.6- and 2.6-fold, respectively). These results show that 4-HNE mediates apoptosis through its effects on JNK and caspase 3, and that 4-HNE metabolizing GST isozyme(s) may be important in the regulation of this pathway of oxidative-stress-induced apoptosis.

Publication types

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

MeSH terms

  • Aldehydes / pharmacology*
  • Apoptosis*
  • Blotting, Northern
  • Blotting, Western
  • Cell Separation
  • Cysteine Proteinase Inhibitors / pharmacology
  • DNA Fragmentation
  • Dose-Response Relationship, Drug
  • Flow Cytometry
  • Glutathione Transferase / chemistry*
  • Glutathione Transferase / isolation & purification
  • Glutathione Transferase / metabolism*
  • HL-60 Cells
  • Humans
  • In Situ Nick-End Labeling
  • Isoenzymes
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinases / metabolism*
  • Oxidative Stress
  • Signal Transduction*
  • Time Factors
  • Transcription Factor AP-1 / metabolism
  • Transfection


  • Aldehydes
  • Cysteine Proteinase Inhibitors
  • Isoenzymes
  • Transcription Factor AP-1
  • Glutathione Transferase
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinases
  • 4-hydroxy-2-nonenal