Failure of zinc to inhibit hyperthermia-induced apoptosis predicts differential nuclease activity detection by comet assay

Autor:	D.W. Fairbairn, Kim L. O'Neill
Rok vydání:	1996
Předmět:	Pharmacology Cancer Research DNA damage Biochemistry (medical) Clinical Biochemistry Apoptotic DNA fragmentation Pharmaceutical Science Cell Biology Cycloheximide Biology Molecular biology Comet assay chemistry.chemical_compound chemistry Cell culture Apoptosis DNA fragmentation DNA
Zdroj:	Apoptosis. 1:81-90
ISSN:	1573-675X 1360-8185
DOI:	10.1007/bf00142082
Popis:	Apoptotic DNA cleavage generally proceeds in two stages, first producing large 50–300kb fragments, and later oligonucleosomal pieces which create the characteristic DNA ladder. We show that zinc treatment of hyperthermia-induced apoptotic cultures is sufficient to prevent ladder formation, but not apoptosis (all features of which were inhibited by actinomycin D and cycloheximide). DNA damage measured in single cells using the comet (single cell gel) assay is detectable in zinc treated cultures using both alkaline and non-denaturing conditions. Both assays predict the same fraction of cells undergoing apoptosis, and damage is detectable earlier than shown by DNA ladder appearance. We conclude that the comet assay is detecting damage consistent with the initial 50–300kb fragments. Additionally, various cell lines when heattreated follow different temporal pathways or display differential apoptotic phenotypes. Also, we were unable to demonstrate an ‘apoptotic window’ for cells refractory to hyperthermia by increasing the heat load.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::65a20a7bcf5b6f7ddfe261de27ce13fc https://doi.org/10.1007/bf00142082 Zobrazit plný text záznamu Full text from SpringerLink