The S-phase cytotoxicity of camptothecin

Abstract

The DNA topoisomerase I inhibitor camptothecin (CAM) is selectively cytotoxic to S-phase cells of HL-60, and some other myelogenous leukemic lines. The early effects of cell exposure to 0.05-0.2 micrograms/ml CAM are seen after 2 h; at that time a progressive degradation of DNA in the chromatin of S-phase cells is initiated. The degradation manifests by "pulverization" of chromatin followed by coalescence of the fine granules and nuclear disintegration. Between 2 and 6 h of treatment, a loss of about 30-70% of DNA from S-phase nuclei is detected by flow cytometry. A 10-min pulse of CAM is adequate to trigger subsequent DNA degradation. Agarose gel electrophoresis of DNA from CAM-treated cells reveals a typical nucleosome core particles "ladder," suggestive of preferential degradation of spacer DNA. Despite extensive loss of DNA and nuclear disintegration, the cell membrane of CAM-treated S-phase cells remains intact for several hours, excluding trypan blue or propidium iodide. Mitochondria, assayed for their ability to maintain a transmembrane potential (rhodamine 123 retention), as well as the lysosomal proton pump (probed by supravital uptake of acridine orange) also remain unchanged in these cells. G1 cells are refractory to CAM under these conditions. Synchronization of cells in S phase by aphidicolin increases the sensitivity of the whole cell population to CAM. The data suggest that CAM or other topoisomerase I inhibitors may be effective in some myelogenous leukemias, especially in combination with treatments synchronizing cells in S phase.