[Radiation-induced DNA fragmentation evaluated by anti-poly(ADP-ribose)immunolabeling in CHO cells. Standardization with pulsed-field electrophoresis].
The poly (ADP-ribose) polymerase is an ubiquitous nuclear protein capable of binding specifically to DNA strand breaks. It synthesizes ADP-ribose polymers proportionally to DNA breaks. The actual method of reference to determine DNA double strand breaks is pulsed-field gel electrophoresis, but this requires many cells. It thus appeared of interest to use poly (ADP-ribos)ylation to follow and estimate gamma-ray-induced DNA fragmentation at the level of isolated cells after gamma-irradiation in chinese hamster ovary cells (CHO-K1). The results obtained by the immunolabelling technique of ADP-ribose polymers were compared to those obtained by pulsed-field gel electrophoresis. They show that poly (ADP-ribos)ylation reflects the occurrence of radiation-induced DNA strand breaks. A clear relationship exists between the amount of ADP-ribose polymers detected and DNA double strand breaks after gamma-irradiation.
Laboratoire d'anatomo-pathologic, hôpital Sainte-Anne, Paris, France.
P
P
Varlet
N
N
Bidon
G
G
Noël
G
G
DeMurcia
J
J
Salamero
D
D
Averbeck
D000818:Animals; D000911:Antibodies, Monoclonal; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000528:radiation effects*; D053938:DNA Fragmentation; D016521:Electrophoresis, Gel, Pulsed-Field; D005455:Fluorescent Antibody Technique; D005720:Gamma Rays; D007118:Immunoassay; D018613:Microscopy, Confocal; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis*
D000911:Antibodies, Monoclonal; D011064:Poly Adenosine Diphosphate Ribose; D004247:DNA
Fragmentation radio-induite de l'ADN évaluée par immunomarquage anti poly (ADP-ribose) dans les CHO. Etalonnage par électrophorèse en champ pulsé.