Increased poly(ADP-ribosyl)ation in intact cells by cisplatin treatment.
Cisplatin (DDP) is a clinically important antitumor drug that induces the formation of DNA-DNA and DNA-protein crosslinks. We have studied whether poly(ADP-ribosyl)ation, a post-translational modification of nuclear proteins that is drastically increased by the presence of DNA strand breaks and plays a role in DNA repair, is induced following DDP treatment of cell cultures. By using an immunofluorescence technique for the in situ detection of poly(ADP-ribose) in intact cells, we found spotty nuclear signals after DDP treatment of O-342 rat ovarian tumor cells or CV-1 monkey cells, but not in untreated control cells, nor in DDP-treated cells postincubated with the ADP-ribosylation inhibitor 3-aminobenzamide. Our results thus provide direct evidence for an involvement of poly(ADP-ribosyl)ation in the cell's response to DDP treatment and, more generally, illustrate the versatility of this rapid in situ method for the detection of increased poly(ADP-ribosyl)ation in living cells.
Bürkle
A
A
DKFZ, Abt. 0610, Heidelberg, Germany.
Chen
G
G
Küpper
J H
JH
Grube
K
K
Zeller
W J
WJ
D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D002945:Cisplatin / Q000494:pharmacology*; D004260:DNA Repair; D004273:DNA, Neoplasm / Q000187:drug effects; D004790:Enzyme Induction / Q000187:drug effects; D005260:Female; D005455:Fluorescent Antibody Technique; D010051:Ovarian Neoplasms / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000096:biosynthesis*; D051381:Rats; D013997:Time Factors; D014407:Tumor Cells, Cultured
D001549:Benzamides; D004273:DNA, Neoplasm; C025160:3-aminobenzamide; D011065:Poly(ADP-ribose) Polymerases; D002945:Cisplatin