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001-es BibID:BIBFORM004656
035-os BibID:WOS:000088961100053
Első szerző:Bacsó Zsolt (biofizikus)
Cím:The DNA of annexin V-binding apoptotic cells is highly fragmented / Bacso, Z., Everson, R. B., Eliason, J. F.
Dátum:2000
Megjegyzések:Jurkat leukemia cells induced to undergo apoptosis by treatment with an antibody against the Fas receptor have two annexin V (AV)-binding subpopulations: (a) single-positive cells that bind AV but not propidium iodide (PI); and (b) double-positive cells that bind AV and PI. The single-positive population is thought to represent an early stage of apoptosis. We have examined the relationship between AV binding and a classical characteristic of apoptosis, DNA fragmentation. Time course studies with Jurkat cells treated for 1, 2, or 4 h with anti-Fas indicated that the proportion of AV-binding cells was increased after 2 h. A significant increase in DNA fragmentation was observed only at 4 h as measured by the mean tail moment determined with the alkaline single cell gel electrophoresis (comet) assay. This correlation suggests a temporal relationship between the two parameters, but does not provide direct evidence of what happens in individual cells. We developed a method to measure fluorescent markers of cellular structure or function with a laser scanning cytometer and then perform the comet assay on the same cells. Cells in each AV-binding subpopulation were re-examined before and after electrophoresis. Most AV-/PI- cells had no DNA damage, although a few cells showed a pattern of damage characteristic for apoptosis. Double-positive cells all had damaged DNA; approximately half had the apoptotic pattern, and the rest had a pattern typical for necrosis. Nearly all of the single-positive cells had damaged DNA with the apoptotic pattern. Both AV-positive populations contained cells with little or no detectable DNA after electrophoresis, indicating that the DNA was highly fragmented. These results indicate that AV binding is an excellent marker for apoptotic cells, but that these cells already have fragmented DNA.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Annexin V
Antibodies
Antibodies, Monoclonal
Antigens, CD95
Apoptosis
Cell Membrane
Cells
Comet Assay
cytology
Dna
DNA Damage
DNA Fragmentation
DNA, Neoplasm
Dyes
Flow Cytometry
Human
immunology
Jurkat Cells
metabolism
Necrosis
pharmacology
Phosphatidylserines
physiology
Propidium
Support, Non-U.S.Gov't
Tumor Cells, Cultured
külföldön készült közlemény
Megjelenés:Cancer research. - 60 : 16 (2000), p. 4623-4628. -
További szerzők:Everson, Richard B. Eliason, James F.
Internet cím:elektronikus változat
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