Application of fluorescence microscopy to measure apoptosis in Jurkat T cells after treatment with a new investigational anticancer agent (N.C.1213).

1. Apoptosis as the mechanism of cell death induced by a new cytotoxic and anticancer agent (N.C.1213) was investigated by morphological and biochemical criteria in human Jurkat T leukemia cells. 2. The effect of N.C.1213 on the survival of Jurkat T, LV-50, H-9, and Molt-3 cells was measured. Jurkat T cells exhibited the highest response, with less than 10% of the cells remaining viable after exposure to 10 microM N.C.1213 for a 24 hr period. All other cell cultures were also affected but to a lesser extent. 3. With the use of a fluorescence microscope, several morphological features characteristic of apoptosis such as condensed chromatin and apoptotic bodies were identified in Jurkat T cells after exposure to N.C.1213 and melphalan. The results indicated that melphalan was more cytotoxic than N.C.1213 as shown by the dye exclusion test. However, N.C.1213 showed a greater apoptotic index than melphalan. The IC50 of N.C.1213 in Jurkat T cells was determined to be 3.5 microM. 4. A DNA ladder (fragmentation of DNA into multimers of approximately 200 base pairs), which is one characteristic feature of apoptosis, was not detected when Jurkat T cells were exposed to N.C.1213. Hence it is probable that the key morphological events in apoptosis observed in the present experimental conditions precede the internucleosomal cleavage of DNA.
AuthorsS C Vashishtha, A J Nazarali, J R Dimmock
JournalCellular and molecular neurobiology (Cell Mol Neurobiol) Vol. 18 Issue 4 Pg. 437-45 (Aug 1998) ISSN: 0272-4340 [Print] UNITED STATES
PMID9619299 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antineoplastic Agents
  • N.C.1213
  • Piperidines
  • Melphalan
  • Antineoplastic Agents (pharmacology)
  • Apoptosis (drug effects)
  • Cell Survival (drug effects)
  • DNA Fragmentation
  • Humans
  • Jurkat Cells (cytology, drug effects, physiology)
  • Kinetics
  • Melphalan (toxicity)
  • Microscopy, Fluorescence (methods)
  • Piperidines (toxicity)
  • Tumor Cells, Cultured

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