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Journal of Cell Science, Vol 107, Issue 9 2483-2489, Copyright © 1994 by Company of Biologists
JOURNAL ARTICLES |
GR Bicknell, RT Snowden and GM Cohen
MRC Toxicology Unit, Leicester University, UK.
Inhibitors of macromolecular synthesis and topoisomerases induce apoptosis in the human leukaemic cell line, U937. In this study, U937 cells were treated with the RNA synthesis inhibitor, actinomycin D (1 microM), the protein synthesis inhibitors, emetine (1 microM) and cycloheximide (100 microM), the topoisomerase II inhibitor, teniposide (5 microM), or the topoisomerase I inhibitor, camptothecin (1 microM). Apoptotic cell death was assessed both by flow cytometry and agarose gel electrophoresis, and was correlated to the appearance of large (20 to > or = 580 kilobase pairs) DNA fragments, as assessed by field inversion gel electrophoresis. In all cases, the appearance of DNA fragments of 20-50 kilobase pairs accompanied the appearance of an apoptotic population and of internucleosomal cleavage. However, teniposide additionally induced a marked increase in fragmentation to > or = 580 kilobase pairs. The cotreatment of cells with zinc (1 mM) inhibited the formation of all large DNA fragments, internucleosomal cleavage and the appearance of an apoptotic population. We conclude that the generation of large DNA fragments is characteristic of apoptosis induced by various stimuli in U937, as has been found previously in rat thymocytes. However, unlike what occurs in rat thymocytes, zinc treatment does not dissociate the formation of large fragments from conventional markers of apoptosis.
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