The role of cell cycle progression in cisplatin-induced apoptosis in Chinese hamster ovary cells.
Many anticancer drugs arrest cells in G2 of the cell cycle and subsequently induce cell death by apoptosis. The current experiments establish a detailed sequence of events that occur in Chinese hamster ovary CHO/UV41 cells following incubation with cisplatin. Synchronized CHO/UV41 cells were damaged with cisplatin in early S. The cells progressed at a normal rate through S but arrested in G2. The arrested cells exhibited normal levels of the mitosis-promoting kinase p34cdc2 in its fully phosphorylated, inactive form. After a protracted arrest, the cells dephosphorylated p34cdc2 and underwent an aberrant mitosis and cytokinesis in which the chromosomes segregated unequally due to the formation of multipolar mitotic spindles. These cells subsequently lost contact with the extracellular matrix, and only then digested their DNA in a manner characteristic of apoptosis. This sequence of events could be dramatically accelerated by the addition of caffeine to G2-arrested cells, which induced dephosphorylation of p34cdc2 and passage through an aberrant mitosis. It has previously been suggested that protein synthesis is required for both caffeine-induced premature mitosis and apoptosis. However, when added in G2, cycloheximide could inhibit neither the caffeine-induced mitosis nor apoptosis. Inhibition was only seen if cycloheximide was added during S before complete synthesis of the proteins required for mitosis. These results demonstrate that, in this model, the proteins thought to be involved in apoptosis are those required for normal cell cycle progression. It is hypothesized that the DNA digestion results from loss of signal transduction originating from the extracellular matrix but that earlier events leading to loss of cell adhesion are critical for the induction of apoptosis.
Demarcq
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Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03755.
Bunch
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RT
Creswell
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Eastman
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D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D016466:CHO Cells / Q000187:drug effects*; D002110:Caffeine / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity*; D006224:Cricetinae; D004247:DNA / Q000187:drug effects; D008938:Mitosis / Q000187:drug effects; D010766:Phosphorylation; D014176:Protein Biosynthesis
D002110:Caffeine; D004247:DNA; D016203:CDC2 Protein Kinase; D002945:Cisplatin