Document Detail

Apoptosis in HL-60 cells induced by 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX).
MedLine Citation:
PMID:  9366896     Owner:  NLM     Status:  MEDLINE    
The potent bacterial mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX), which is formed during chlorination of drinking water, has been studied with respect to induction of cell death in promyelocytic leukemic HL-60 cells. Cells exposed to MX for 1 h and further incubated for 3 h, revealed no significant increase in the proportion of cells with compromised plasma membrane damage as judged by trypan blue or propidium iodide exclusion. However, flow cytometric studies and microscopic analysis of HL-60 cells after staining with Giemsa or Hoechst 33342, revealed that more than 30% of the cells exposed to 30-100 microM of MX, showed the characteristic morphology and biochemical markers of apoptosis. On the other hand, in cultures exposed to 300 microM MX, less than 5% of the cells appeared to be apoptotic (< G1 DNA) 3 h after treatment, which is similar to control values. Microscopic analysis of Hoechst 33342-stained cells revealed that they were 'arrested' in the early stages of chromatin condensation, but these cells eventually became necrotic. Some decrease in the percentage of cells in S-phase was observed 3 h after exposure to MX (10, 30 and 100 microM), but the induced cell death was not markedly cell stage specific. The characteristic ladder pattern of apoptotic cells was observed when DNA isolated from MX-exposed HL-60 cells was electrophoresed in agarose. The apoptotic process could also be detected by analysis with alkaline filter elution (AE), as a decrease in the total DNA recovered; and by single cell gel electrophoresis, as a decrease in the average number of cells/comets observable on each slide. With the protocols used no apparent increase in values in the normalized area above the curve (NAAC) (alkaline elution) or tail moments (single cell gel electrophoresis (SCGE)) were detected, indicating that apoptotic cells are not necessarily a confounding factor when assaying for genotoxicity with these techniques.
U Marsteinstredet; R Wiger; G Brunborg; J K Hongslo; J A Holme
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Chemico-biological interactions     Volume:  106     ISSN:  0009-2797     ISO Abbreviation:  Chem. Biol. Interact.     Publication Date:  1997 Sep 
Date Detail:
Created Date:  1997-12-08     Completed Date:  1997-12-08     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0227276     Medline TA:  Chem Biol Interact     Country:  IRELAND    
Other Details:
Languages:  eng     Pagination:  89-107     Citation Subset:  IM    
Department of Environmental Medicine, National Institute of Public Health, Oslo, Norway.
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MeSH Terms
Apoptosis / drug effects*
Azure Stains
Cell Cycle / drug effects,  physiology
Cell Membrane / drug effects
Cell Size / drug effects
Cell Survival / drug effects
DNA Damage
DNA Fragmentation / drug effects
Electrophoresis, Agar Gel
Flow Cytometry
Furans / toxicity*
HL-60 Cells
Mutagens / toxicity*
Propidium / metabolism
Trypan Blue / metabolism
Reg. No./Substance:
0/Azure Stains; 0/Benzimidazoles; 0/Furans; 0/Mutagens; 23491-52-3/HOE 33342; 36015-30-2/Propidium; 72-57-1/Trypan Blue; 77439-76-0/3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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