Alkaline elution analysis of DNA fragmentation induced during apoptosis.
We report that alkaline elution analysis fails to provide a complete profile of DNA fragmentation induced by some genotoxic agents, particularly if these agents induce apoptotic cell death resulting in fragmentation of DNA into oligonucleosomal length multimers. It was questioned whether these oligonucleosome fragments are responsible for the more rapidly eluting component of DNA that is observed as a biphasic elution profile when cells treated with certain genotoxic agents are subjected to alkaline elution. The results of this study indicate that DNA fragmented during apoptotic cell death is eluted in fractions before alkaline denaturation that are normally discarded. Collection of fractions prior to alkaline denaturation is recommended for complete evaluation of the total spectrum of damage induced by genotoxic agents, especially when the compound is suspected of inducing cell death by apoptosis.
Division of Laboratory Research, AMC Cancer Research Center, Denver, Colorado 80214.
M R
MR
Kaeck
S
S
Briggs
H J
HJ
Thompson
D000468:Alkalies; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D004247:DNA / Q000187:drug effects / Q000302:isolation & purification*; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000187:drug effects / Q000302:isolation & purification; D004277:DNA, Single-Stranded / Q000187:drug effects / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D007939:Leukemia L1210 / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009691:Nucleic Acid Denaturation; D012643:Selenium / Q000494:pharmacology; D018038:Sodium Selenite; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000473:pathology
D000468:Alkalies; D004273:DNA, Neoplasm; D004277:DNA, Single-Stranded; D004247:DNA; D012643:Selenium; D018038:Sodium Selenite