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Taxol-Induced Mitotic Block Triggers Rapid Onset of a p53-Independent Apoptotic Pathway

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Abstract

Background

At therapeutic concentrations, the antineoplastic agent taxol selectively perturbs mitotic spindle microtubules. Taxol has recently been shown to induce apoptosis, similar to the mechanism of cell death induced by other antineoplastic agents. However, taxol has shown efficacy against drug-refractory cancers, raising the possibility that this pharmacological agent may trigger an alternative apoptotic pathway.

Materials and Methods

The kinetics and IC50 of mitotic (M) block, aberrant mitosis, and cytotoxicity following taxol treatment were analyzed in human cell lines as well as normal mouse embryo fibroblasts (MEFs) and MEFs derived from p53-null mice. Apoptosis was followed by DNA gel electrophoresis and by in situ DNA end-labeling (TUNEL).

Results

Taxol induced two forms of cell cycle arrest: either directly in early M at prophase or, for those cells progressing through aberrant mitosis, arrest in G1 as multimininucleated cells. TUNEL labeling revealed that DNA nicking occurred within 30 min of the arrest in prophase. In contrast, G1-arrested, multimininucleated cells became TUNEL positive only after several days. In the subset of cells that became blocked directly in prophase, both wt p53-expressing and p53-null MEFs responded similarly to taxol, showing rapid onset of DNA nicking and apoptosis. However, p53-null MEFs progressing through aberrant mitosis failed to arrest in the subsequent G1 phase or to become TUNEL positive, and remained viable.

Conclusions

Taxol induces two forms of cell cycle arrest, which in turn induce two independent apoptotic pathways. Arrest in prophase induces rapid onset of a p53-independent pathway, whereas G1-block and the resulting slow (3–5 days) apoptotic pathway are p53 dependent.

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Acknowledgments

We greatly appreciate the assistance of Judy Miller in carrying out FACS analysis and also thank J. Xu for helpful discussions on the TUNEL method and Dr. Helen Ranney for valuable input during the completion of this manuscript.

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  1. Catherine M. Woods

    Present address: Alliance Pharmaceutical Co., 3040 Science Park Road, San Diego, CA, 92121, USA

  2. Elias Lazarides

    Present address: Astral Inc., 3040 Science Park Road, San Diego, CA, 92121, USA

Authors and Affiliations

  1. Department of Pharmacology, Merck Research Laboratories, West Point, Pennsylvania, USA

    Catherine M. Woods, Jian Zhu, Patricia A. McQueney, Daniel Bollag & Elias Lazarides

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Woods, C.M., Zhu, J., McQueney, P.A. et al. Taxol-Induced Mitotic Block Triggers Rapid Onset of a p53-Independent Apoptotic Pathway. Mol Med 1, 506–526 (1995). https://doi.org/10.1007/BF03401588

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