Inactivation of the mitotic checkpoint as a determinant of the efficacy of microtubule-targeted drugs in killing human cancer cells.

Drugs that disrupt microtubule dynamics include some of the most important of cancer chemotherapies. While these drugs, which include paclitaxel (Taxol), are known to invoke the mitotic checkpoint, the factors that determine cancer cell killing remain incompletely characterized. Cells that are relatively resistant to killing by these drugs block robustly in mitosis, whereas cells sensitive to killing block only transiently in mitosis before undergoing nuclear fragmentation and death. Passage through mitosis was an absolute requirement of drug-induced death, because death was markedly reduced in cells blocked at both G(1)-S and G(2). Cell killing was at least in part linked to the absence or inactivation of BubR1, a kinetochore-associated phosphoprotein that mediates the mitotic checkpoint. Sensitivity to paclitaxel correlated with decreased BubR1 protein expression in human cancer cell lines, including those derived from breast and ovarian cancers. Silencing of BubR1 via RNA interference inactivated the mitotic checkpoint in drug-resistant cells, and reversed resistance to paclitaxel and nocodazole. Together, these results suggest that the mitotic checkpoint is an important determinant of the efficacy of microtubule-targeting drugs in killing cancer cells, potentially providing novel targets for increasing treatment efficacy.

[1]  M. Cammer,et al.  Gene expression and mitotic exit induced by microtubule-stabilizing drugs. , 2003, Cancer research.

[2]  Takashi Takahashi,et al.  Association between mitotic spindle checkpoint impairment and susceptibility to the induction of apoptosis by anti-microtubule agents in human lung cancers. , 2003, The American journal of pathology.

[3]  Z. Bonday,et al.  Centromere-associated protein-E is essential for the mammalian mitotic checkpoint to prevent aneuploidy due to single chromosome loss , 2003, The Journal of cell biology.

[4]  Y. Mao,et al.  Activating and Silencing the Mitotic Checkpoint through CENP-E-Dependent Activation/Inactivation of BubR1 , 2003, Cell.

[5]  K. Sullivan,et al.  Suppression of centromere dynamics by Taxol in living osteosarcoma cells. , 2003, Cancer research.

[6]  T. Ried,et al.  Inhibition of BUB1 results in genomic instability and anchorage-independent growth of normal human fibroblasts. , 2003, Cancer research.

[7]  Stephen S. Taylor,et al.  Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores , 2003, The Journal of cell biology.

[8]  M. Guenther,et al.  Histone deacetylase 4 interacts with 53BP1 to mediate the DNA damage response , 2003, The Journal of cell biology.

[9]  A. Riva,et al.  Second generation taxanes: from the natural framework to the challenge of drug resistance. , 2003, Current medicinal chemistry. Anti-cancer agents.

[10]  K. Sullivan,et al.  Centromeres and Kinetochores From Epigenetics to Mitotic Checkpoint Signaling , 2003, Cell.

[11]  A. Musacchio,et al.  The spindle checkpoint: structural insights into dynamic signalling , 2002, Nature Reviews Molecular Cell Biology.

[12]  Rey-Huei Chen,et al.  BubR1 is essential for kinetochore localization of other spindle checkpoint proteins and its phosphorylation requires Mad1 , 2002, The Journal of cell biology.

[13]  E. Gabrielson,et al.  Variable levels of chromosomal instability and mitotic spindle checkpoint defects in breast cancer. , 2002, The American journal of pathology.

[14]  S. Kaufmann,et al.  G1 and G2 cell-cycle arrest following microtubule depolymerization in human breast cancer cells , 2002 .

[15]  E. Salmon,et al.  Inducing precocious anaphase in cultured mammalian cells. , 2002, Cell motility and the cytoskeleton.

[16]  Stephen S. Taylor,et al.  Kinetochore localisation and phosphorylation of the mitotic checkpoint components Bub1 and BubR1 are differentially regulated by spindle events in human cells. , 2001, Journal of cell science.

[17]  B. Hill,et al.  Mechanism of mitotic block and inhibition of cell proliferation by the semisynthetic Vinca alkaloids vinorelbine and its newer derivative vinflunine. , 2001, Molecular pharmacology.

[18]  W. Mckenna,et al.  Detection of repair activity during the DNA damage-induced G2 delay in human cancer cells , 2001, Oncogene.

[19]  Carl W. Miller,et al.  Mutations in the mitotic check point gene, MAD1L1, in human cancers , 2001, Oncogene.

[20]  M. Kavallaris,et al.  The role of β-tubulin isotypes in resistance to antimitotic drugs , 2001 .

[21]  B. Sikic,et al.  MDR 1 activation is the predominant resistance mechanism selected by vinblastine in MES-SA cells , 2000, British Journal of Cancer.

[22]  E. Nogales,et al.  A common pharmacophore for epothilone and taxanes: molecular basis for drug resistance conferred by tubulin mutations in human cancer cells. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[23]  L. Milas,et al.  Effect of docetaxel on the therapeutic ratio of fractionated radiotherapy in vivo. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[24]  T. Fojo,et al.  Molecular effects of paclitaxel: Myths and reality (a critical review) , 1999, International journal of cancer.

[25]  G. Chan,et al.  Human Bubr1 Is a Mitotic Checkpoint Kinase That Monitors Cenp-E Functions at Kinetochores and Binds the Cyclosome/APC , 1999, The Journal of cell biology.

[26]  L. Meijer,et al.  Paullones, a series of cyclin-dependent kinase inhibitors: synthesis, evaluation of CDK1/cyclin B inhibition, and in vitro antitumor activity. , 1999, Journal of medicinal chemistry.

[27]  P Wadsworth,et al.  Taxol suppresses dynamics of individual microtubules in living human tumor cells. , 1999, Molecular biology of the cell.

[28]  M. Polymeropoulos,et al.  Cytogenetic and molecular characterization of random chromosomal rearrangements activating the drug resistance gene, MDR1/P‐glycoprotein, in drug‐selected cell lines and patients with drug refractory ALL , 1998, Genes, chromosomes & cancer.

[29]  L. Milas,et al.  Docetaxel enhances tumor radioresponse in vivo. , 1997, Clinical cancer research : an official journal of the American Association for Cancer Research.

[30]  M. Kavallaris,et al.  Taxol-resistant epithelial ovarian tumors are associated with altered expression of specific beta-tubulin isotypes. , 1997, The Journal of clinical investigation.

[31]  R. Muschel,et al.  Cyclin B1 availability is a rate-limiting component of the radiation-induced G2 delay in HeLa cells. , 1997, Cancer research.

[32]  C. Runowicz,et al.  Analysis of MDR1 expression in normal and malignant endometrium by reverse transcription-polymerase chain reaction and immunohistochemistry. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[33]  R. Benjamin,et al.  Phase II study of paclitaxel in patients with previously treated osteosarcoma and its variants , 1996, Cancer.

[34]  J. Lynch,et al.  A phase II trial of paclitaxel in the treatment of recurrent or metastatic soft tissue sarcomas or bone sarcomas. , 1996, Journal of experimental therapeutics & oncology.

[35]  J. B. Rattner,et al.  CENP-F is a protein of the nuclear matrix that assembles onto kinetochores at late G2 and is rapidly degraded after mitosis , 1995, The Journal of cell biology.

[36]  A. C. Dubbelman,et al.  Pharmacokinetics of paclitaxel and metabolites in a randomized comparative study in platinum-pretreated ovarian cancer patients. , 1993, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[37]  M. Jordan,et al.  Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[38]  M. Jordan,et al.  Effects of vinblastine, podophyllotoxin and nocodazole on mitotic spindles. Implications for the role of microtubule dynamics in mitosis. , 1992, Journal of cell science.

[39]  M. Gottesman,et al.  Isolation of a taxol-resistant Chinese hamster ovary cell mutant that has an alteration in alpha-tubulin. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[40]  J. Lee,et al.  Effects of nocodazole on structures of calf brain tubulin. , 1980, Biochemistry.

[41]  P. Schiff,et al.  Taxol stabilizes microtubules in mouse fibroblast cells. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[42]  M. De Brabander,et al.  Interaction of oncodazole (R 17934), a new antitumoral drug, with rat brain tubulin. , 1976, Biochemical and biophysical research communications.

[43]  G. Chan,et al.  The mitotic checkpoint: a signaling pathway that allows a single unattached kinetochore to inhibit mitotic exit. , 2003, Progress in cell cycle research.

[44]  S. Kaufmann,et al.  G(1) and G(2) cell-cycle arrest following microtubule depolymerization in human breast cancer cells. , 2002, The Journal of clinical investigation.

[45]  M. Shichiri,et al.  Genetic and epigenetic inactivation of mitotic checkpoint genes hBUB1 and hBUBR1 and their relationship to survival. , 2002, Cancer research.

[46]  Erich A. Nigg,et al.  Cell division: Mitotic kinases as regulators of cell division and its checkpoints , 2001, Nature Reviews Molecular Cell Biology.

[47]  B. Pollock.  Obstacles and opportunities for the use of health‐related quality‐of‐life assessment in pediatric cancer clinical trials (discussion) , 1999, International journal of cancer. Supplement = Journal international du cancer. Supplement.