Hj. Steinfelder et al., A fast and sensitive technique to study the kinetics and the concentrationdependencies of DNA fragmentation during drug-induced apoptosis, J PHARM TOX, 43(1), 2000, pp. 79-84
Citations number
15
Categorie Soggetti
Pharmacology & Toxicology
Journal title
JOURNAL OF PHARMACOLOGICAL AND TOXICOLOGICAL METHODS
Apoptotic cell death with its characteristic coordinated cellular breakdown
can be triggered by cytotoxic drugs. One prominent feature that differenti
ates apoptotic from necrotic cell death is the caspase-mediated activation
of an endonuclease that internucleosomally cleaves DNA resulting in the so-
called apoptotic DNA ladder. Here we report a new rapid, sensitive and inex
pensive column separation technique to study drug-induced DNA fragmentation
from 10(6) or less cells. This technique, which is based on a modified pla
smid spin column kit, avoids the use of hazardous chemicals. With this proc
edure and subsequent densitometric analysis it was possible to study the co
ncentration dependencies and the kinetics of drug-induced DNA fragmentation
.. The applicability of this technique is shown for okadaic-acid- and canth
aridic-acid-treated pituitary GH(3) cells as well as highly okadaic acid-re
sistant sublines. These studies allowed us to compare as well as to differe
ntiate the effects and potencies of these structurally different but functi
onally quite similar inhibitors of ser/thr phosphatases 1 and 2A. (C) 2000
Elsevier Science Inc. All rights reserved.