Poly(ADP-ribose) synthesis: a useful parameter for identifying apoptotic cells.
Histochem J
; 29(11-12): 831-7, 1997.
Article
em En
| MEDLINE
| ID: mdl-9466150
ABSTRACT
Cell death by apoptosis was analysed in HeLa cells either treated with the antitumoral drug bleomycin or depleted of growth factors by long-term culture without medium change. The interference of apoptosis with normal cell cycle progression was followed by flow cytometry in cells stained with propidium iodide and with antibody to S-phase-related PCNA protein. Bleomycin-treated cells showed a net accumulation in G2/M phase paralleled by the appearance of material with a subdiploid DNA content. Cells with a subdiploid DNA content were also present in growth factor-depleted cultures and were shown to derive from all the cell cycle phases. To identify apoptotic features in HeLa cell cultures, we applied a recently developed assay based on the simultaneous analysis in the single cell of three parameters, namely chromatin condensation, DNA degradation and poly(ADP-ribose) synthesis. Apoptotic cells were visualized by sequential reactions Hoechst staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling assay and immunoreaction with anti-poly(ADP-ribose) monoclonal antibody. Positive reactions were obtained for cells at different stages of the apoptotic programme showing condensed nuclei, fragmented chromatin and apoptotic bodies.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Poli Adenosina Difosfato Ribose
/
Apoptose
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Histochem J
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Itália