Docetaxel induces cell death through mitotic catastrophe in human breast cancer cells

Apoptosis has long been considered to be the prevailing mechanism of cell death in response to chemotherapy. Currently, a more heterogeneous model of tumor response to therapy is acknowledged wherein multiple modes of death combine to generate the overall tumor response. The resulting mechanisms of cell death are likely determined by the mechanism of action of the drug, the dosing regimen used, and the genetic background of the cells within the tumor. This study describes a nonapoptotic response to docetaxel therapy in human breast cancer cells of increasing cancer progression (MCF-10A, MCF-7, and MDA-mb-231). Docetaxel is a microtubule-stabilizing taxane that is being used in the clinic for the treatment of breast and prostate cancers and small cell carcinoma of the lung. The genetic backgrounds of these cells were characterized for the status of key pathways and gene products involved in drug response and cell death. Cellular responses to docetaxel were assessed by characterizing cell viability, cell cycle checkpoint arrest, and mechanisms of cell death. Mechanisms of cell death were determined by Annexin V binding and scoring of cytology-stained cells by morphology and transmission electron microscopy. The primary mechanism of death was determined to be mitotic catastrophe by scoring of micronucleated cells and cells undergoing aberrant mitosis. Other, nonapoptotic modes of death were also determined. No significant changes in levels of apoptosis were observed in response to docetaxel.

[1]  A. Klein-Szanto,et al.  Transformation of Human Breast Epithelial Cells by c‐Ha‐ras Oncogene , 1991, Molecular carcinogenesis.

[2]  M. Sarker,et al.  Stimulation of the mitogen-activated protein kinase pathway antagonizes TRAIL-induced apoptosis downstream of BID cleavage in human breast cancer MCF-7 cells , 2002, Oncogene.

[3]  R. Burghardt,et al.  Rb localization and phosphorylation kinetics correlate with the cellular phenotype of cultured breast adenocarcinoma cells , 2002, In Vitro Cellular & Developmental Biology - Animal.

[4]  U. Göhring,et al.  Lack of correlation between P53 expression, BCL-2 expression, apoptosis and ex vivo chemosensitivity in advanced human breast cancer. , 2000, Anticancer research.

[5]  J. Crown,et al.  The taxanes: an update , 2000, The Lancet.

[6]  P. Vrignaud,et al.  Preclinical evaluation of docetaxel (Taxotere). , 1995, Seminars in oncology.

[7]  S. Kaufmann,et al.  A multistep model for paclitaxel-induced apoptosis in human breast cancer cell lines. , 2001, Experimental cell research.

[8]  W. B. Derry,et al.  Mitotic block induced in HeLa cells by low concentrations of paclitaxel (Taxol) results in abnormal mitotic exit and apoptotic cell death. , 1996, Cancer research.

[9]  R. Dickson,et al.  Adhesion-regulated G1 cell cycle arrest in epithelial cells requires the downregulation of c-Myc , 2001, Oncogene.

[10]  Korey R. Johnson,et al.  In vitro evaluation of schedule-dependent interactions between docetaxel and doxorubicin against human breast and ovarian cancer cells. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[11]  A. Giaccia,et al.  Loss of p21Waf1/Cip1 sensitizes tumors to radiation by an apoptosis-independent mechanism. , 1997, Cancer research.

[12]  C. Thompson,et al.  Mitochondrial membrane potential regulates matrix configuration and cytochrome c release during apoptosis , 2003, Cell Death and Differentiation.

[13]  G. Chaudhuri,et al.  Nitric oxide-induced cytostasis and cell cycle arrest of a human breast cancer cell line (MDA-MB-231): Potential role of cyclin D1 , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[14]  S. McLeskey,et al.  Quantitative demonstration of spontaneous metastasis by MCF-7 human breast cancer cells cotransfected with fibroblast growth factor 4 and LacZ. , 1993, Cancer research.

[15]  S. Moir,et al.  Conjugated linoleic acids (CLAs) regulate the expression of key apoptotic genes in human breast cancer cells , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  D. Fabbro,et al.  bcl‐xl antisense treatment induces apoptosis in breast carcinoma cells , 2000, International journal of cancer.

[17]  J. Chovancová,et al.  Radiation-induced apoptosis and cell cycle alterations in human carcinoma cell lines with different radiosensitivities. , 2000, Neoplasma.

[18]  E. Chang,et al.  Inhibitory effects of the combination of HER-2 antisense oligonucleotide and chemotherapeutic agents used for the treatment of human breast cancer , 2001, Cancer Gene Therapy.

[19]  K. Kohn,et al.  Abrogation of an S-phase checkpoint and potentiation of camptothecin cytotoxicity by 7-hydroxystaurosporine (UCN-01) in human cancer cell lines, possibly influenced by p53 function. , 1997, Cancer research.

[20]  J. Little,et al.  Absence of a radiation-induced first-cycle G1-S arrest in p53+ human tumor cells synchronized by mitotic selection. , 1998, Cancer research.

[21]  Qin Wang,et al.  All-trans retinoic acid potentiates Taxotere-induced cell death mediated by Jun N-terminal kinase in breast cancer cells , 2004, Oncogene.

[22]  A. Long,et al.  A human cell line from a pleural effusion derived from a breast carcinoma. , 1973, Journal of the National Cancer Institute.

[23]  D. Gewirtz.  Growth arrest and cell death in the breast tumor cell in response to ionizing radiation and chemotherapeutic agents which induce DNA damage , 2000, Breast Cancer Research and Treatment.

[24]  C. Elferink,et al.  Progressive resistance to apoptosis in a cell lineage model of human proliferative breast disease. , 2001, Journal of the National Cancer Institute.

[25]  T. Thomas,et al.  Induction of p21 (CIP1/WAF1/SID1) by estradiol in a breast epithelial cell line transfected with the recombinant estrogen receptor gene: , 2004, Breast Cancer Research and Treatment.

[26]  Bernhard O. Palsson,et al.  Cancer cell lines , 1999 .

[27]  C. Croce,et al.  Bcl2 is the guardian of microtubule integrity. , 1997, Cancer research.

[28]  K. Geisinger,et al.  nm23—Relationship to the metastatic potential of breast carcinoma cell lines, primary human xenografts, and lymph node negative breast carcinoma patients , 1997, Cancer.

[29]  H. Iwase,et al.  [Breast cancer]. , 2006, Nihon rinsho. Japanese journal of clinical medicine.

[30]  T. Jacks,et al.  Characterization of the p53-Dependent Postmitotic Checkpoint following Spindle Disruption , 1998, Molecular and Cellular Biology.

[31]  H. Kawai,et al.  Direct interaction between BRCA1 and the estrogen receptor regulates vascular endothelial growth factor (VEGF) transcription and secretion in breast cancer cells , 2002, Oncogene.

[32]  A. Eastman,et al.  A novel indolocarbazole, ICP-1, abrogates DNA damage-induced cell cycle arrest and enhances cytotoxicity: similarities and differences to the cell cycle checkpoint abrogator UCN-01. , 2002, Molecular cancer therapeutics.

[33]  Lianfeng Zhang,et al.  Role of Raf-1 and FAK in cell density-dependent regulation of integrin-dependent activation of MAP kinase. , 2002, Carcinogenesis.

[34]  B. Wouters,et al.  Apoptosis, p53, and tumor cell sensitivity to anticancer agents. , 1999, Cancer research.

[35]  E. Bernhard,et al.  How does radiation kill cells? , 1999, Current opinion in chemical biology.

[36]  E. Kandel,et al.  A senescence-like phenotype distinguishes tumor cells that undergo terminal proliferation arrest after exposure to anticancer agents. , 1999, Cancer research.

[37]  T. K. Yeung,et al.  The mode of action of taxol: apoptosis at low concentration and necrosis at high concentration. , 1999, Biochemical and biophysical research communications.

[38]  M. Olivé,et al.  Breast tumor cell lines from pleural effusions. , 1974, Journal of the National Cancer Institute.

[39]  Longgui Wang,et al.  The effect of antimicrotubule agents on signal transduction pathways of apoptosis: a review , 1999, Cancer Chemotherapy and Pharmacology.

[40]  R. Gillies,et al.  Determination of cell number in monolayer cultures. , 1986, Analytical biochemistry.

[41]  R. Lock,et al.  Dual modes of death induced by etoposide in human epithelial tumor cells allow Bcl-2 to inhibit apoptosis without affecting clonogenic survival. , 1996, Cancer research.

[42]  I. Roninson,et al.  If not apoptosis, then what? Treatment-induced senescence and mitotic catastrophe in tumor cells. , 2001, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[43]  B. Jeffy,et al.  Disruption of cell cycle kinetics by benzo[a]pyrene: inverse expression patterns of BRCA-1 and p53 in MCF-7 cells arrested in S and G2. , 2000, Neoplasia.

[44]  M. Shibata,et al.  Apoptotic detection methods--from morphology to gene. , 2003, Progress in histochemistry and cytochemistry.

[45]  N. Watanabe,et al.  The presence of single nucleotide instability in human breast cancer cell lines. , 2001, Cancer research.

[46]  C Haanen,et al.  A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. , 1995, Journal of immunological methods.

[47]  F. Sarkar,et al.  Differential sensitivity of normal and malignant breast epithelial cells to genistein is partly mediated by p21(WAF1). , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[48]  S. Horwitz,et al.  Mechanisms of Taxol-induced cell death are concentration dependent. , 1998, Cancer research.

[49]  T. Fojo,et al.  Loss of cell cycle control allows selective microtubule-active drug-induced Bcl-2 phosphorylation and cytotoxicity in autonomous cancer cells. , 2000, Cancer research.

[50]  C. Park,et al.  Lipotropes regulate BCL-2 gene expression in the human breast cancer cell line, MCF-7 , 2002, In Vitro Cellular & Developmental Biology - Animal.

[51]  R. Müller,et al.  Crosstalk of the mitotic spindle assembly checkpoint with p53 to prevent polyploidy , 2004, Oncogene.

[52]  C. Chi,et al.  Comparison of 2‐methoxyestradiol‐induced, docetaxel‐induced, and paclitaxel‐induced apoptosis in hepatoma cells and its correlation with reactive oxygen species , 2000, Cancer.

[53]  V. Heinemann,et al.  Gemcitabine: metabolism, mechanisms of action, and self-potentiation. , 1995, Seminars in oncology.

[54]  Y. Pommier,et al.  Apoptotic response to camptothecin and 7‐hydroxystaurosporine (UCN‐01) in the 8 human breast cancer cell lines of the NCI anticancer drug screen: Multifactorial relationships with topoisomerase i, protein kinase C, Bcl‐2, p53, MDM‐2 and caspase pathways , 1999, International Journal of Cancer.

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

[56]  W D Lawrence,et al.  Paclitaxel‐induced apoptosis in MCF‐7 breast‐cancer cells , 1997, International journal of cancer.

[57]  D. Noh,et al.  Functional screening of genes suppressing TRAIL-induced apoptosis: distinct inhibitory activities of Bcl-XL and Bcl-2 , 2003, British Journal of Cancer.

[58]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.

[59]  H. Pinedo,et al.  Variation in the kinetics of caspase-3 activation, Bcl-2 phosphorylation and apoptotic morphology in unselected human ovarian cancer cell lines as a response to docetaxel. , 2002, Biochemical pharmacology.

[60]  J. Díaz,et al.  Assembly of purified GDP-tubulin into microtubules induced by taxol and taxotere: reversibility, ligand stoichiometry, and competition. , 1993, Biochemistry.

[61]  I. Ringel,et al.  Studies with RP 56976 (taxotere): a semisynthetic analogue of taxol. , 1991, Journal of the National Cancer Institute.

[62]  E. Radany,et al.  erbB-2 overexpression in human mammary epithelial cells confers growth factor independence. , 1999, Endocrinology.

[63]  T. Fojo,et al.  Paclitaxel-Induced FasL-Independent Apoptosis and Slow (Non-Apoptotic) Cell Death , 2002, Cancer biology & therapy.

[64]  L. Milas,et al.  Lack of correlation between mitotic arrest or apoptosis and antitumor effect of docetaxel , 1999, Cancer Chemotherapy and Pharmacology.

[65]  Human Cell Culture , 2002 .

[66]  A. Agarwal,et al.  Epidermal growth factor promotes MDA-MB-231 breast cancer cell migration through a phosphatidylinositol 3'-kinase and phospholipase C-dependent mechanism. , 1999, Cancer research.

[67]  Y. Yoo,et al.  Novel bile acid derivatives induce apoptosis via a p53-independent pathway in human breast carcinoma cells. , 2001, Cancer letters.

[68]  A. Suzuki,et al.  Necessity of interleukin-1beta converting enzyme cascade in taxotere-initiated death signaling. , 1998, European journal of pharmacology.