The DNA of annexin V-binding apoptotic cells is highly fragmented

Z. Bacsó, R. B. Everson, J. F. Eliason

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish
Pages (from-to)4623-4628
Number of pages6
JournalCancer Research
Volume60
Issue number16
Publication statusPublished - Aug 15 2000

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Annexin A5
DNA
Comet Assay
Propidium
Apoptosis
Jurkat Cells
DNA Fragmentation
Electrophoresis
CD95 Antigens
Cellular Structures
Population
DNA Damage
Leukemia
Lasers
Necrosis

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Bacsó, Z., Everson, R. B., & Eliason, J. F. (2000). The DNA of annexin V-binding apoptotic cells is highly fragmented. Cancer Research, 60(16), 4623-4628.

The DNA of annexin V-binding apoptotic cells is highly fragmented. / Bacsó, Z.; Everson, R. B.; Eliason, J. F.

In: Cancer Research, Vol. 60, No. 16, 15.08.2000, p. 4623-4628.

Research output: Contribution to journalArticle

Bacsó, Z, Everson, RB & Eliason, JF 2000, 'The DNA of annexin V-binding apoptotic cells is highly fragmented', Cancer Research, vol. 60, no. 16, pp. 4623-4628.
Bacsó Z, Everson RB, Eliason JF. The DNA of annexin V-binding apoptotic cells is highly fragmented. Cancer Research. 2000 Aug 15;60(16):4623-4628.
Bacsó, Z. ; Everson, R. B. ; Eliason, J. F. / The DNA of annexin V-binding apoptotic cells is highly fragmented. In: Cancer Research. 2000 ; Vol. 60, No. 16. pp. 4623-4628.
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