Mitotic block induced in HeLa cells by low concentrations of paclitaxel (Taxol) results in abnormal mitotic exit and apoptotic cell death.
Paclitaxel at low concentrations (10 nM for 20 h) induces approximately 90% mitotic block at the metaphase/anaphase transition in HeLa cells, apparently by suppressing dynamics of spindle microtubules (M. A. Jordan et al., Proc. Natl. Acad. Sci. USA, 90: 9552-9556, 1993). It is not known, however, whether inhibition of mitosis by such low paclitaxel concentrations results in cell death. In the present work, we found that after removal of paclitaxel (10 nM-1 microM), blocked cells did not resume proliferation. Instead, cells exited mitosis abnormally within 24 h. They did not progress through anaphase or cytokinesis but entered an interphase-like state (chromatin decondensed, and an interphase-like microtubule array and nuclear membranes reformed). Many cells (> or = 55%) contained multiple nuclei. Additional DNA synthesis and polyploidy did not occur. DNA degradation into nucleosome-sized fragments characteristic of apoptosis began during drug incubation and increased after drug removal. Cells died within 48-72 h. Incubation with paclitaxel (10 nM for 20 h) resulted in high intracellular drug accumulation (8.3 microM) and little efflux after paclitaxel removal; intracellular retention of paclitaxel may contribute to its efficacy. The results support the hypothesis that the most potent chemotherapeutic mechanism of paclitaxel is kinetic stabilization of spindle microtubule dynamics.
Jordan
M A
MA
Department of Molecular, Cellular, and Developmental Biology, University of California Santa Barbara 93106, USA.
Wendell
K
K
Gardiner
S
S
Derry
W B
WB
Copp
H
H
Wilson
L
L
D000705:Anaphase; D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism / Q000633:toxicity*; D001692:Biological Transport; D002453:Cell Cycle / Q000187:drug effects*; D016923:Cell Death / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002843:Chromatin / Q000187:drug effects / Q000648:ultrastructure; D002877:Chromosomes, Human / Q000187:drug effects / Q000648:ultrastructure; D004273:DNA, Neoplasm / Q000187:drug effects / Q000302:isolation & purification; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008677:Metaphase; D008870:Microtubules / Q000187:drug effects / Q000648:ultrastructure; D008938:Mitosis / Q000187:drug effects; D017239:Paclitaxel / Q000378:metabolism / Q000633:toxicity*; D013997:Time Factors
D000972:Antineoplastic Agents, Phytogenic; D002843:Chromatin; D004273:DNA, Neoplasm; D017239:Paclitaxel