Linked Life Data

1333943

Source:http://linkedlifedata.com/resource/pubmed/id/1333943

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
1993-1-13
pubmed:abstractText
The present review describes several methods to characterize and differentiate between two different mechanisms of cell death, apoptosis and necrosis. Most of these methods were applied to studies of apoptosis triggered in the human leukemic HL-60 cell line by DNA topoisomerase I or II inhibitors, and in rat thymocytes by either topoisomerase inhibitors or prednisolone. In most cases, apoptosis was selective to cells in a particular phase of the cell cycle: only S-phase HL-60 cells and G0 thymocytes were mainly affected. Necrosis was induced by excessively high concentrations of these drugs. The following cell features were found useful to characterize the mode of cell death: a) Activation of an endonuclease in apoptocic cells resulted in extraction of the low molecular weight DNA following cell permeabilization, which, in turn, led to their decreased stainability with DNA-specific fluorochromes. Measurements of DNA content made it possible to identify apoptotic cells and to recognize the cell cycle phase specificity of the apoptotic process. b) Plasma membrane integrity, which is lost in necrotic but not apoptotic cells, was probed by the exclusion of propidium iodide (PI). The combination of PI followed by Hoechst 33342 proved to be an excellent probe to distinguish live, necrotic, early- and late-apoptotic cells. c) Mitochondrial transmembrane potential, assayed by retention of rhodamine 123 was preserved in apoptotic but not necrotic cells. d) The ATP-dependent lysosomal proton pump, tested by the supravital uptake of acridine orange (AO) was also preserved in apoptotic but not necrotic cells. e) Bivariate analysis of cells stained for DNA and protein revealed markedly diminished protein content in apoptotic cells, most likely due to activation of endogenous proteases. Necrotic cells, having leaky membranes, had minimal protein content. f) Staining of RNA allowed for the discrimination of G0 from G1 cells and thus made it possible to reveal that apoptosis was selective to G0 thymocytes. g) The decrease in forward light scatter, paralleled either by no change (HL-60 cells) or an increase (thymocytes) of right angle scatter, were early changes during apoptosis. h) The sensitivity of DNA in situ to denaturation, was increased in apoptotic and necrotic cells. This feature, probed by staining with AO at low pH, provided a sensitive and early assay to discriminate between live, apoptotic and necrotic cells, and to evaluate the cell cycle phase specificity of these processes. i) The in situ nick translation assay employing labeled triphosphonucleotides can be used to reveal DNA strand breaks, to detect the very early stages of apoptosis.(ABSTRACT TRUNCATED AT 400 WORDS)
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
0196-4763
pubmed:author
pubmed:issnType
Print
pubmed:volume
13
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
795-808
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:1333943-Animals, pubmed-meshheading:1333943-Antineoplastic Agents, pubmed-meshheading:1333943-Apoptosis, pubmed-meshheading:1333943-Cell Cycle, pubmed-meshheading:1333943-Cell Death, pubmed-meshheading:1333943-Cell Membrane, pubmed-meshheading:1333943-Cell Size, pubmed-meshheading:1333943-DNA, pubmed-meshheading:1333943-DNA Damage, pubmed-meshheading:1333943-Endodeoxyribonucleases, pubmed-meshheading:1333943-Endopeptidases, pubmed-meshheading:1333943-Enzyme Activation, pubmed-meshheading:1333943-Flow Cytometry, pubmed-meshheading:1333943-Fluorescent Dyes, pubmed-meshheading:1333943-Humans, pubmed-meshheading:1333943-Leukemia, Promyelocytic, Acute, pubmed-meshheading:1333943-Membrane Potentials, pubmed-meshheading:1333943-Nucleic Acid Denaturation, pubmed-meshheading:1333943-Organelles, pubmed-meshheading:1333943-Proteins, pubmed-meshheading:1333943-Rats, pubmed-meshheading:1333943-Thymus Gland, pubmed-meshheading:1333943-Topoisomerase I Inhibitors, pubmed-meshheading:1333943-Topoisomerase II Inhibitors, pubmed-meshheading:1333943-Tumor Cells, Cultured
pubmed:year
1992
pubmed:articleTitle
Features of apoptotic cells measured by flow cytometry.
pubmed:affiliation
Cancer Research Institute, New York Medical College, Valhalla 10595.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Review, Research Support, Non-U.S. Gov't