title: PARP degradation in apoptotic Syrian hamster embryo (SHE) cells compared to HL60 cell line.
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.
authors: EBSE-Centre des Sciences de l'Environnement, Université de Metz, Faculté des Sciences, rue Delestraint, 57070, Metz, France.
authors: S
authors: S
authors: Alexandre
authors: C
authors: C
authors: Rast
authors: G
authors: G
authors: Nguyen-Ba
authors: G G
authors: GG
authors: Poirier
authors: P
authors: P
authors: Vasseur
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D053938:DNA Fragmentation; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism*; D005047:Etoposide / Q000494:pharmacology; D018922:HL-60 Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008647:Mesocricetus; D009243:NAD / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000276:immunology / Q000378:metabolism*
chemical_list: D016895:Culture Media, Serum-Free; D009243:NAD; D005047:Etoposide; D011065:Poly(ADP-ribose) Polymerases