Apoptosis of rat thymocytes triggered by prednisolone, camptothecin, or teniposide is selective to G0 cells and is prevented by inhibitors of proteases.
Rat thymocytes were treated in culture with prednisolone or the DNA topoisomerase I or II inhibitors, camptothecin (CAM) or teniposide (TN), and proportions of cells in different phases of the cell cycle were estimated by flow cytometry using a staining methodology which makes it possible to discriminate between G0 and G1 cells, as well as to recognize the cells which undergo apoptosis. The appearance of apoptotic cells in cultures treated with pharmacological concentrations of these drugs, observed as early as 3-6 hr after treatment, coincided with the selective loss of G0 cells in these cultures, while no significant changes in the proportion of S or G2+M cells were apparent. Agarose gel electrophoresis of DNA isolated from the treated cells indicated degradation of the internucleosomal spacer sections, typical of the endonucleolytic activity which accompanies apoptotic cell death. The data indicate that G0 thymocytes were particularly sensitive to agents that induce apoptosis while cells progressing through the cell cycle were resistant. This suggests that under in vivo conditions (immunological response), the selective death of G0 cells may promote the clonal expansion of stimulated thymocytes which enter the cell cycle. Together with our earlier studies on the effects of CAM and TN on MOLT-4 and HL-60 leukemic cell lines, these data indicate that both, phenotypic- and and cell cycle phase specific- factors modify the ability of cells to respond to toxic agents, including chemotherapeutics by apoptosis.(ABSTRACT TRUNCATED AT 250 WORDS)
Bruno
S
S
Cancer Research Institute, New York Medical College, Valhalla 10523.
Lassota
P
P
Giaretti
W
W
Darzynkiewicz
Z
Z
D000818:Animals; D002166:Camptothecin / Q000633:toxicity*; D002453:Cell Cycle; D016923:Cell Death / Q000187:drug effects*; D004247:DNA / Q000737:chemistry; D005434:Flow Cytometry; D066298:In Vitro Techniques; D011239:Prednisolone / Q000633:toxicity*; D011480:Protease Inhibitors / Q000494:pharmacology*; D012313:RNA / Q000737:chemistry; D051381:Rats; D013713:Teniposide / Q000633:toxicity*; D013950:Thymus Gland / Q000166:cytology*; D059004:Topoisomerase I Inhibitors
D011480:Protease Inhibitors; D059004:Topoisomerase I Inhibitors; D012313:RNA; D004247:DNA; D013713:Teniposide; D011239:Prednisolone; D002166:Camptothecin