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Nuclear changes in necrotic HL-60 cells.

R Bortul1, M Zweyer, A M Billi

  • 1Dipartimento di Morfologia Umana Normale, Università di Trieste, 34138 Trieste, Italy.

Journal of Cellular Biochemistry. Supplement
|July 17, 2001
PubMed
Summary
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Necrotic cell death involves distinct nuclear changes, including protein cleavage and altered nuclear matrix morphology, differentiating it from apoptosis. These findings provide new criteria for distinguishing between these cell death pathways.

Area of Science:

  • Cell biology
  • Molecular biology
  • Biochemistry

Background:

  • Cell death in eukaryotes occurs via apoptosis or necrosis.
  • Apoptosis involves defined nuclear changes like proteolysis and DNA fragmentation.
  • Nuclear modifications during necrosis are less understood.

Purpose of the Study:

  • Investigate nuclear modifications during ethanol-induced necrotic cell death in HL-60 cells.
  • Determine if nuclear protein changes during necrosis differ from apoptosis.
  • Identify biochemical and morphological markers to distinguish necrosis from apoptosis.

Main Methods:

  • Immunofluorescence staining of nuclear proteins (lamin B1, NuMA, topoisomerase IIalpha, SC-35, B23/nucleophosmin, C23/nucleolin, UBF, fibrillarin, RNA polymerase I).
  • Immunoblotting analysis to detect protein cleavage.

Related Experiment Videos

  • Transmission electron microscopy of nuclear matrix fractions.
  • Treatment with caspase inhibitors.

    • Main Results:

    • Immunofluorescence showed altered spatial distribution of some nuclear proteins (NuMA, lamin B1, topoisomerase IIalpha, SC-35, B23/nucleophosmin) during necrosis.
    • NuMA redistribution resembled apoptotic changes, while others remained unchanged (C23/nucleolin, UBF, fibrillarin, RNA polymerase I).
    • Nuclear proteins SAF-A, SATB1, and NuMA were cleaved during necrosis; SATB1 cleavage mirrored apoptosis.
    • Lamin B1 and topoisomerase IIalpha were not cleaved in necrosis but were in apoptosis.
    • Transmission electron microscopy revealed slight nuclear matrix rarefaction in necrotic cells, less severe than in apoptotic cells.
    • Caspase inhibitors blocked apoptosis but not necrosis-induced protein cleavage.

    Conclusions:

    • Necrotic cell death induces significant biochemical and morphological changes at the nuclear level.
    • These nuclear alterations are distinct from those observed in apoptosis.
    • Identified novel biochemical and morphological criteria to differentiate between necrotic and apoptotic cell death.