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.
Vashishtha
S C
SC
Medicinal Chemistry Research Laboratory, College of Pharmacy and Nutrition, University of Saskatchewan, Saskatoon, Canada.
Nazarali
A J
AJ
Dimmock
J R
JR
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation; D006801:Humans; D019169:Jurkat Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D007700:Kinetics; D008558:Melphalan / Q000633:toxicity*; D008856:Microscopy, Fluorescence / Q000379:methods; D010880:Piperidines / Q000633:toxicity*; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; C113730:N.C.1213; D010880:Piperidines; D008558:Melphalan