Mitotic Spindle Disruption by Alternating Electric Fields Leads to Improper Chromosome Segregation and Mitotic Catastrophe in Cancer Cells

Tumor Treating Fields (TTFields) are low intensity, intermediate frequency, alternating electric fields. TTFields are a unique anti-mitotic treatment modality delivered in a continuous, noninvasive manner to the region of a tumor. It was previously postulated that by exerting directional forces on highly polar intracellular elements during mitosis, TTFields could disrupt the normal assembly of spindle microtubules. However there is limited evidence directly linking TTFields to an effect on microtubules. Here we report that TTFields decrease the ratio between polymerized and total tubulin, and prevent proper mitotic spindle assembly. The aberrant mitotic events induced by TTFields lead to abnormal chromosome segregation, cellular multinucleation, and caspase dependent apoptosis of daughter cells. The effect of TTFields on cell viability and clonogenic survival substantially depends upon the cell division rate. We show that by extending the duration of exposure to TTFields, slowly dividing cells can be affected to a similar extent as rapidly dividing cells.

[1]  C. Bassett.  The development and application of pulsed electromagnetic fields (PEMFs) for ununited fractures and arthrodeses. , 1985, Clinics in plastic surgery.

[2]  Robert C. Wolpert,et al.  A Review of the , 1985 .

[3]  A. Hyman,et al.  Real-time visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts , 1990, Cell.

[4]  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.

[5]  D E Ingber,et al.  Extracellular matrix controls tubulin monomer levels in hepatocytes by regulating protein turnover. , 1994, Molecular biology of the cell.

[6]  M. Jordan,et al.  Microtubule dynamics: taking aim at a moving target. , 1995, Chemistry & biology.

[7]  E. Lazarides,et al.  Epothilones, a new class of microtubule-stabilizing agents with a taxol-like mechanism of action. , 1995, Cancer research.

[8]  S. Schreiber,et al.  Understanding and controlling the cell cycle with natural products. , 1996, Chemistry & biology.

[9]  P. Duesberg,et al.  Aneuploidy correlated 100% with chemical transformation of Chinese hamster cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Hyman,et al.  Coupling cell division and cell death to microtubule dynamics. , 1997, Current opinion in cell biology.

[11]  A Elhajouji,et al.  Spindle poisons can induce polyploidy by mitotic slippage and micronucleate mononucleates in the cytokinesis-block assay. , 1998, Mutagenesis.

[12]  K. Zilles,et al.  Comparison of iodotyrosines and methionine uptake in a rat glioma model. , 1998, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[13]  P. Nisen,et al.  Taxol, vincristine or nocodazole induces lethality in G1-checkpoint-defective human astrocytoma U373MG cells by triggering hyperploid progression. , 1999, Carcinogenesis.

[14]  B. Sikic,et al.  Mechanisms of action of and resistance to antitubulin agents: microtubule dynamics, drug transport, and cell death. , 1999, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[15]  E. Elson,et al.  Biologic Effects of Radiofrequency and Microwave Fields: In Vivo and In Vitro Experimental Results , 1999 .

[16]  B. Leone,et al.  Vinorelbine and paclitaxel as first-line chemotherapy in metastatic breast cancer. , 1999, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[17]  M. Repacholi,et al.  Interaction of static and extremely low frequency electric and magnetic fields with living systems: health effects and research needs. , 1999, Bioelectromagnetics.

[18]  E. Sahai,et al.  Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I , 2001, Nature Cell Biology.

[19]  D. Compton,et al.  Chromosome Movement in Mitosis Requires Microtubule Anchorage at Spindle Poles , 2001, The Journal of cell biology.

[20]  Richard J. Cebula,et al.  A Cost Benefit Analysis of Voting , 2002 .

[21]  Jean Marx,et al.  Debate Surges Over the Origins of Genomic Defects in Cancer , 2002, Science.

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

[23]  E. Parry,et al.  Detection and characterization of mechanisms of action of aneugenic chemicals. , 2002, Mutagenesis.

[24]  Choong-Ryoul Sihn,et al.  p55CDC/hCDC20 mutant induces mitotic catastrophe by inhibiting the MAD2-dependent spindle checkpoint activity in tumor cells. , 2003, Cancer letters.

[25]  M. Samii,et al.  Morphometrical Characterization of Two Glioma Models in the Brain of Immunocompetent and Immunodeficient Rats , 1999, Journal of Neuro-Oncology.

[26]  E. Dekel,et al.  Disruption of cancer cell replication by alternating electric fields. , 2004, Cancer research.

[27]  M. Jordan,et al.  Microtubules as a target for anticancer drugs , 2004, Nature Reviews Cancer.

[28]  Helder Maiato,et al.  Stuck in division or passing through: what happens when cells cannot satisfy the spindle assembly checkpoint. , 2004, Developmental cell.

[29]  C. Walczak,et al.  Mitotic spindle assembly and chromosome segregation: refocusing on microtubule dynamics. , 2004, Molecular cell.

[30]  A. Mercurio,et al.  Hypoxia stimulates carcinoma invasion by stabilizing microtubules and promoting the Rab11 trafficking of the alpha6beta4 integrin. , 2005, Cancer research.

[31]  B. Weaver,et al.  Decoding the links between mitosis, cancer, and chemotherapy: The mitotic checkpoint, adaptation, and cell death. , 2005, Cancer cell.

[32]  Min Zhao,et al.  Controlling cell behavior electrically: current views and future potential. , 2005, Physiological reviews.

[33]  T. Chow,et al.  DNA repair protein: endo-exonuclease as a new frontier in cancer therapy. , 2005, Future oncology.

[34]  G. Gorbsky,et al.  Spindle checkpoint function and cellular sensitivity to antimitotic drugs , 2006, Molecular Cancer Therapeutics.

[35]  S. Noguchi.  Predictive factors for response to docetaxel in human breast cancers , 2006, Cancer science.

[36]  E. Salmon,et al.  Measuring nanometer scale gradients in spindle microtubule dynamics using model convolution microscopy. , 2006, Molecular biology of the cell.

[37]  E. Salmon,et al.  The spindle-assembly checkpoint in space and time , 2007, Nature Reviews Molecular Cell Biology.

[38]  E. Dekel,et al.  Alternating electric fields arrest cell proliferation in animal tumor models and human brain tumors , 2007, Proceedings of the National Academy of Sciences.

[39]  Cristina Montagna,et al.  Aneuploidy acts both oncogenically and as a tumor suppressor. , 2007, Cancer cell.

[40]  C. Caldas,et al.  The Extracellular Matrix Protein TGFBI Induces Microtubule Stabilization and Sensitizes Ovarian Cancers to Paclitaxel , 2007, Cancer cell.

[41]  Gerard H Markx,et al.  The use of electric fields in tissue engineering , 2008, Organogenesis.

[42]  M. Blagosklonny,et al.  Mechanism of G1-like arrest by low concentrations of paclitaxel: next cell cycle p53-dependent arrest with sub G1 DNA content mediated by prolonged mitosis , 2008, Oncogene.

[43]  T. Mitchison,et al.  Cell type variation in responses to antimitotic drugs that target microtubules and kinesin-5. , 2008, Cancer research.

[44]  Th. Boveri.  Concerning the Origin of Malignant Tumours by Theodor Boveri. Translated and annotated by Henry Harris , 2008, Journal of Cell Science.

[45]  Yangzhong Tang,et al.  Quantitative live imaging of cancer and normal cells treated with Kinesin-5 inhibitors indicates significant differences in phenotypic responses and cell fate , 2008, Molecular Cancer Therapeutics.

[46]  Stephen S. Taylor,et al.  How do anti-mitotic drugs kill cancer cells? , 2009, Journal of Cell Science.

[47]  D. Cleveland,et al.  Boveri revisited: chromosomal instability, aneuploidy and tumorigenesis , 2009, Nature Reviews Molecular Cell Biology.

[48]  T. Mitchison,et al.  Evidence that mitotic exit is a better cancer therapeutic target than spindle assembly. , 2009, Cancer cell.

[49]  J. Vymazal,et al.  Chemotherapeutic treatment efficacy and sensitivity are increased by adjuvant alternating electric fields (TTFields) , 2009, BMC medical physics.

[50]  S. Badar,et al.  Potent inhibition of tubulin polymerisation and proliferation of paclitaxel-resistant 1A9PTX22 human ovarian cancer cells by albendazole. , 2009, Anticancer research.

[51]  Michal Cifra,et al.  Electric field generated by axial longitudinal vibration modes of microtubule , 2010, Biosyst..

[52]  Linjie Guo,et al.  The ATM–p53 pathway suppresses aneuploidy-induced tumorigenesis , 2010, Proceedings of the National Academy of Sciences.

[53]  Duane A. Compton,et al.  Proliferation of aneuploid human cells is limited by a p53-dependent mechanism , 2010, The Journal of cell biology.

[54]  M. Cifra,et al.  High-frequency electric field and radiation characteristics of cellular microtubule network. , 2011, Journal of theoretical biology.

[55]  L. Galluzzi,et al.  Mitotic catastrophe: a mechanism for avoiding genomic instability , 2011, Nature Reviews Molecular Cell Biology.

[56]  P. Gutin,et al.  NovoTTF-100A versus physician's choice chemotherapy in recurrent glioblastoma: a randomised phase III trial of a novel treatment modality. , 2012, European journal of cancer.

[57]  Thomas H. Lee,et al.  Taking AIM at a Moving Target , 2012 .

[58]  J. Baehring,et al.  Faculty Opinions recommendation of NovoTTF-100A versus physician's choice chemotherapy in recurrent glioblastoma: a randomised phase III trial of a novel treatment modality. , 2012 .

[59]  Hailing Yang,et al.  The Role of Microtubules and Their Dynamics in Cell Migration* , 2012, The Journal of Biological Chemistry.

[60]  Til Aach,et al.  Signal and Noise Modeling in Confocal Laser Scanning Fluorescence Microscopy , 2012, MICCAI.

[61]  D. Bouscary,et al.  Flow Cytometric Quantification of All Phases of the Cell Cycle and Apoptosis in a Two-Color Fluorescence Plot , 2013, PloS one.

[62]  Yoram Palti,et al.  Tumor treating fields: a new frontier in cancer therapy , 2013, Annals of the New York Academy of Sciences.

[63]  Lauren M. Zasadil,et al.  Chromosome missegregation rate predicts whether aneuploidy will promote or suppress tumors , 2013, Proceedings of the National Academy of Sciences.

[64]  Yun-chi Tang,et al.  Gene Copy-Number Alterations: A Cost-Benefit Analysis , 2013, Cell.

[65]  Miklos Pless,et al.  A phase I/II trial of Tumor Treating Fields (TTFields) therapy in combination with pemetrexed for advanced non-small cell lung cancer. , 2013, Lung cancer.

[66]  S. Toms,et al.  The effect of field strength on glioblastoma multiforme response in patients treated with the NovoTTF™-100A system , 2014, World Journal of Surgical Oncology.

[67]  Shuvomoy Banerjee,et al.  Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Can Induce Chromosomal Instability through Targeted Degradation of the Mitotic Checkpoint Kinase Bub1 , 2014, Journal of Virology.

[68]  N. Butowski,et al.  Clinical practice experience with NovoTTF-100A™ system for glioblastoma: The Patient Registry Dataset (PRiDe). , 2014, Seminars in oncology.

[69]  Moshe Giladi,et al.  Alternating electric fields (tumor-treating fields therapy) can improve chemotherapy treatment efficacy in non-small cell lung cancer both in vitro and in vivo. , 2014, Seminars in oncology.

[70]  Mark E. Burkard,et al.  Cytotoxicity of Paclitaxel in Breast Cancer Is due to Chromosome Missegregation on Multipolar Spindles , 2014, Science Translational Medicine.

[71]  H. Maiato,et al.  N-terminus-modified Hec1 suppresses tumour growth by interfering with kinetochore–microtubule dynamics , 2014, Oncogene.

[72]  P. Basser,et al.  Predicting the electric field distribution in the brain for the treatment of glioblastoma , 2014, Physics in medicine and biology.

[73]  E. Kirson,et al.  Mitotic disruption and reduced clonogenicity of pancreatic cancer cells in vitro and in vivo by tumor treating fields. , 2014, Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.].

[74]  E. Wong,et al.  An Evidence-Based Review of Alternating Electric Fields Therapy for Malignant Gliomas , 2015, Current Treatment Options in Oncology.