Transfection with 4-hydroxynonenal-metabolizing glutathione S-transferase isozymes leads to phenotypic transformation and immortalization of adherent cells

Eur J Biochem. 2004 May;271(9):1690-701. doi: 10.1111/j.1432-1033.2004.04067.x.


4-Hydroxy-2-trans-nonenal (4-HNE), one of the major end products of lipid peroxidation, has been shown to induce apoptosis in a variety of cell lines. It appears to modulate signaling processes in more than one way because it has been suggested to have a role in signaling for differentiation and proliferation. We show for the first time that incorporation of 4-HNE-metabolizing glutathione S-transferase (GST) isozyme, hGSTA4-4, into adherent cell lines HLE B-3 and CCL-75, by either cDNA transfection or microinjection of active enzyme, leads to their transformation. The dramatic phenotypic changes due to the incorporation of hGSTA4-4 include rounding of cells and anchorage-independent rapid proliferation of immortalized, rounded, and smaller cells. Incorporation of the inactive mutant of hGSTA4-4 (Y212F) in cells by either microinjection or transfection does not cause transformation, suggesting that the activity of hGSTA4-4 toward 4-HNE is required for transformation. This is further confirmed by the fact that mouse and Drosophila GST isozymes (mGSTA4-4 and DmGSTD1-1), which have high activity toward 4-HNE and subsequent depletion of 4-HNE, cause transformation whereas human GST isozymes hGSTP1-1 and hGSTA1-1, with minimal activity toward 4-HNE, do not cause transformation. In cells overexpressing active hGSTA4-4, expression of transforming growth factor beta1, cyclin-dependent kinase 2, protein kinase C betaII and extracellular signal regulated kinase is upregulated, whereas expression of p53 is downregulated. These studies suggest that alterations in 4-HNE homeostasis can profoundly affect cell-cycle signaling events.

Publication types

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

MeSH terms

  • Aldehydes / metabolism*
  • Bacterial Proteins*
  • Carrier Proteins / physiology*
  • Cell Cycle / genetics
  • Cell Division
  • Cell Line, Transformed
  • Cells, Cultured
  • Glutathione / metabolism
  • Glutathione Transferase / physiology*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Isoenzymes / physiology*
  • JNK Mitogen-Activated Protein Kinases
  • Microinjections
  • Mitogen-Activated Protein Kinases / metabolism
  • Transfection


  • Aldehydes
  • Bacterial Proteins
  • Carrier Proteins
  • GstA protein, bacteria
  • Intracellular Signaling Peptides and Proteins
  • Isoenzymes
  • GSTA1 protein, human
  • Glutathione Transferase
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinases
  • Glutathione
  • 4-hydroxy-2-nonenal