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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1993-3-31
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pubmed:abstractText |
To analyse the putative role of methylation of cytosine residues in the nuclear DNA as a regulatory step during cellular ageing, we incubated ageing human amniotic fluid derived fibroblast-like cells and non-ageing NIH-3T3 cells with 5-azacytidine. BrdUrd/Hoechst and acridine orange (AO) flow cytometry was used to compare the effects of the base analogue on cell proliferation and cell differentiation. In NIH-3T3 cultures, 96h exposures to 4 microM 5-azacytidine caused diminished cell proliferation due to cell arrest in the G1 compartments of the second and third cell cycles of serum stimulated cells. The exit from the G0/G1 compartment was not affected. The 5-azacytidine induced cell kinetic disturbances were unstable in NIH-3T3 cultures, such that pre-treated cells reverted to normal cell cycle transit within 2-3 days after termination of treatment. In contrast, 5-azacytidine pre-treated amniotic fluid derived fibroblast-like cell cultures showed persistently elevated G2 phase arrests and delayed G0/G1 phase exit kinetics, which explain the premature cessation of proliferation observed in these primary cultures. In both cell systems, 5-azacytidine exposed cultures showed elevated numbers of G1 phase cells with increased RNA content as revealed by AO flow cytometry. Again, this effect was reversible in NIH-3T3 cells but not in amniotic fluid derived fibroblast-like cells. These contrasting responses to 5-azacytidine are likely to reflect intrinsic differences in methylation patterns or de novo methylase activity between ageing cell strains and non-ageing cell lines.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0960-7722
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
26
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
45-54
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:7679929-3T3 Cells,
pubmed-meshheading:7679929-Acridine Orange,
pubmed-meshheading:7679929-Amniotic Fluid,
pubmed-meshheading:7679929-Animals,
pubmed-meshheading:7679929-Azacitidine,
pubmed-meshheading:7679929-Bromodeoxyuridine,
pubmed-meshheading:7679929-Cell Aging,
pubmed-meshheading:7679929-Cell Division,
pubmed-meshheading:7679929-Cell Separation,
pubmed-meshheading:7679929-Cells, Cultured,
pubmed-meshheading:7679929-Flow Cytometry,
pubmed-meshheading:7679929-G1 Phase,
pubmed-meshheading:7679929-G2 Phase,
pubmed-meshheading:7679929-Humans,
pubmed-meshheading:7679929-Methylation,
pubmed-meshheading:7679929-Mice,
pubmed-meshheading:7679929-RNA
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pubmed:year |
1993
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pubmed:articleTitle |
Induction of replicative senescence by 5-azacytidine: fundamental cell kinetic differences between human diploid fibroblasts and NIH-3T3 cells.
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pubmed:affiliation |
Department of Human Genetics, University of Würzburg, Germany.
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pubmed:publicationType |
Journal Article,
Comparative Study
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