PARP degradation in apoptotic Syrian hamster embryo (SHE) cells compared to HL60 cell line

Biochimie. 2000 Dec;82(12):1115-22. doi: 10.1016/s0300-9084(00)01194-9.

Abstract

In this study, we attempted to identify apoptotic Syrian hamster embryo (SHE) cells by detecting the specific cleavage of poly(ADP-ribose)polymerase (PARP). Apoptosis was unequivocally identified in serum-deprived SHE cells. After protein electrophoresis and transfer, the anti-PARP antibody (C-2-10) was applied in order to visualize PARP degradation and the anti-polymer antibody (LP96-10) was used to identify PARP and its expected 89-kDa fragment on the membrane after renaturation and NAD+ addition. Results showed that PARP rapidly disappeared during apoptosis in SHE cells, but the resulting fragment remained undetectable with the anti-PARP antibody and no stable polymerase activity of this fragment was measured using anti-polymer antibody. Serum-starved SHE cells were compared to the etoposide-treated HL60 cell line as a control for typical apoptosis-related PARP cleavage. These results underline the fact that while PARP degradation is a criterion for apoptosis, the diagnosis of apoptosis can not rely exclusively on the appearance of its 89-kDa fragment as this signal may fail to appear in some cell systems.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • Blotting, Western
  • Cell Line
  • Cricetinae
  • Culture Media, Serum-Free
  • DNA Fragmentation
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / metabolism*
  • Etoposide / pharmacology
  • HL-60 Cells / drug effects
  • HL-60 Cells / metabolism*
  • Humans
  • Mesocricetus
  • NAD / pharmacology
  • Poly(ADP-ribose) Polymerases / immunology
  • Poly(ADP-ribose) Polymerases / metabolism*

Substances

  • Culture Media, Serum-Free
  • NAD
  • Etoposide
  • Poly(ADP-ribose) Polymerases