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. 1998 Aug;18(4):437–445. doi: 10.1023/A:1022505700642

Application of Fluorescence Microscopy to Measure Apoptosis in Jurkat T Cells After Treatment with a New Investigational Anticancer Agent (N.C.1213)

S C Vashishtha 1, A J Nazarali 2, J R Dimmock 1
PMCID: PMC11560149  PMID: 9619299

Abstract

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 μM 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 indemnified 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 μM

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 internucleosomalcleavage of DNA.

Keywords: apoptosis, human Jurkat T cells, Mannich base, melphalan, fluorescence microscopy

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