Adhesive Interactions in Normal and Transformed Cells
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Yury A. Rovensky Adhesive Interactions in Normal and Transformed Cells
Yury A. Rovensky, M.D., Ph.D., D.Sci. Former Leading Researcher at Cancer Research Center of the Russian Academy of Medical Sciences Moscow, Russia yuryrovenskiy@sbcglobal.net ISBN 978-1-61779-303-5 e-isbn 978-1-61779-304-2 DOI 10.1007/978-1-61779-304-2 Springer New York Dordrecht Heidelberg London Library of Congress Control Number: 2011934257 Springer Science+Business Media, LLC 2011 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. Printed on acid-free paper Humana Press is part of Springer Science+Business Media (www.springer.com)
To my wife Tanya with love
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Preface The ability of tissue cells to be attached to each other and to the surrounding solid substance (extracellular matrix) is a pivotal regulator of major cellular functions such as proliferation, responses to growth-stimulating factors, cell survival, differentiation, and migration of cells in an organism. Therefore, the cellular adhesive interactions play a critical role in basic biological processes such as formation of tissues and organs in embryonic development, maintenance of structural integrity of all tissues in an adult organism, and tissue regeneration and remodeling. The adhesive interactions are also involved in inflammation and degeneration processes, which are at the basis of many diseases. As a result of oncogenic transformation, the adhesive interactions of transformed cells are significantly altered. In the pathological behavior of malignant tumor cells, significant weakening of their ability to adhere to each other, to normal cells, and to the extracellular matrix, plays a key role. Alterations in these adhesive interactions form the basis of invasion and metastasis of malignant tumors. Therefore, the understanding of mechanisms of cellular adhesive interactions and their alterations in malignant tumors is very important in both biological and medical aspects. Adhesive Interactions in Normal and Transformed Cells starts with the description of molecular composition of the extracellular matrix, which tissue cells adhere to. The matrix proteins that are bound with the specific cell surface receptors resulting in the cell-matrix adhesion are also discussed. Several sections are devoted to the cytoskeleton systems. Particular attention is given to the actin filaments and microtubules that play a pivotal role in cell-extracellular matrix and cell cell adhesive interactions, and also in cell migration. The formation, regulation, and dynamics of these cytoskeleton systems are examined. vii
viii Preface Different types of pseudopodia that are formed and used by cells as driving organs during cell spreading and cell migration are described. Various types of specific adhesion structures formed by cells in order to attach to the extracellular matrix are considered. Attention is given to focal adhesions (focal contacts), to their structure, regulation, and dynamics, which play a critical role in cell migration. Several sections are devoted to the intracellular signal transduction pathways. The signaling pathways are triggered by the extracellular molecules (ligands) that bind to specialized cell surface receptor proteins. Different types of cell surface receptors are characterized. Particular attention is given to integrin receptors, which as components of focal adhesions play a key role in cell-matrix attachment and also fulfill functions of transducers of intracellular signals. Different integrin receptormediated signaling pathways that determine and control cell morphology, proliferation, survival, and locomotion are considered. Also, the growth factor receptor-mediated mitogenic and morphogenic signaling pathways are examined. Special attention is given to significant alterations in the integrin mediated cellmatrix adhesion caused by oncogenic transformation of the cells. The consequences of these alterations manifested in such typical traits of transformed cells as weakening of the cell-matrix adhesion, anchorage independence, constitutive mitogenic activation, escape from anoikis, and high locomotory activity are considered. The movement of fibroblastic cells and different factors involved in the cell locomotion machinery are considered. These factors include actin cytoskeleton reorganizations and microtubule dynamics, the phenomenon of contact inhibition of cell locomotion, and dynamic regulation of focal adhesions during cell locomotion. The morphogenic action of soluble growth factors resulting in cell locomotion is also examined. Several sections are devoted to fundamental alterations in cell locomotion machinery caused by oncogenic transformation of the cells. These alterations apply to the pseudopodial activity and focal adhesion formation in transformed cells, and also their sensitivity to growth factors. The ability of cells to respond to the adhesion heterogeneity or various geometrical configurations (topography) of the extracellular matrix surfaces is discussed in detail. The topographic cell responses to cylindrical surfaces of high curvatures or the surface reliefs of various kinds (such as nanoscale or microscale linear grooves, holes, or vertical rods) are examined. These responses apply to the cell shape, locomotion, and other cellular functions. The mechanisms of these cell responses are discussed. The alterations in the topographic cell responses caused by oncogenic transformation of cells are considered. In particular, alterations of the cell shape, changes in the direction of cell migration, and alterations in the functional activities as a result of oncogenic transformation are described. Last chapter of the book is devoted to the intercellular adhesive interactions. The compositions of several types of the intercellular adhesion structures are described. Particular attention is paid to the adherens junctions, their structure and dynamic regulation, which is the basis of cell rearrangement and tissue integrity maintenance.
Preface ix A critical contribution of cadherin receptors and local actin cytoskeleton to the regulation of cell cell adhesion is examined. Signaling pathways coupling cadherinmediated intercellular contacts to cell proliferation are considered. The cell cell adhesion alterations caused by oncogenic transformation of the cells are further examined. These alterations result in uncontrolled proliferation of malignant tumor cells, their inability to form orderly tissue structures, cancer invasion, and metastasis. Adhesive Interactions in Normal and Transformed Cells is based on modern scientific data and includes the results of the author s long-term research. It is intended for researchers, postdocs, undergraduate, and graduate students, whose scientific interests are in the fields of cell biology, cancer biology, cancer research, and developmental biology. West Hollywood, CA Yury A. Rovensky
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Acknowledgements I am extremely grateful to Dr. Alexander V. Ljubimov (Cedars-Sinai Medical Center, Los Angeles, CA) for critical reading of the manuscript, exceptionally helpful comments, stimulating discussions, and valuable advices. Generous help of Julia Y. Moers at all stages of the manuscript preparation including processing of the figures and assistance in preparing the References is highly appreciated. I thank Dr. Eugene B. Mechetner (Stonsa Biopharm, Inc., Irvine, CA) for his friendly encouragement and support. I would like to express my gratitude to Dr.Tatyana M. Svitkina (University of Pennsylvania, Philadelphia, PA) for providing her spectacular TEM photos and her valuable comments, and also to all my collaborators, who gave me their figures; they are acknowledged in the legends. I thank Raymond T. Moers for his help. I am much obliged to all my colleagues and friends, with whom I worked for many years. I apologize to all those whose valuable work in this field have not been referenced due to space limits. xi
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Contents 1 Introduction... 1 References... 5 2 The Extracellular Matrix... 7 References... 10 3 Cytoskeleton... 13 3.1 Actin Filaments... 13 3.1.1 Actin-Binding Proteins... 16 3.1.2 Actin Filament Dynamics... 20 3.2 Microtubules... 24 3.2.1 Motor Proteins... 27 3.2.2 Nonmotor Proteins... 28 3.3 Intermediate Filaments... 29 References... 31 4 Pseudopodia and Adhesion Structures... 37 4.1 The Formation of Pseudopodia... 37 4.2 Cell Extracellular Matrix Adhesion Structures... 44 4.2.1 Focal Contacts (Focal Adhesions)... 45 4.2.2 Focal Complexes... 50 4.2.3 Fibrillar Adhesions... 51 4.2.4 Hemidesmosomes... 51 4.2.5 Podosomes and Invadopodia... 51 References... 53 5 Adhesive Interactions of Tissue Cells with the Extracellular Matrix... 57 5.1 Cell Spreading on the Extracellular Matrix Surface... 57 5.1.1 Cells in a Suspended State... 57 5.1.2 The Morphology of Cell Spreading Process in Normal Cells... 64 xiii
xiv Contents 5.1.3 Morphological Alterations in the Spreading of Transformed Cells... 74 5.2 The Signaling Pathways in the Spread Cells... 88 5.2.1 Cell Surface Receptors... 88 5.2.2 Intracellular Signal Transduction... 91 5.3 Signaling Pathways from Integrin and Growth Factor Receptors in Normal Cells... 94 5.3.1 Integrin Receptor-Mediated Mitogenic Signaling Pathways... 95 5.3.2 Growth Factor Receptor-Mediated Mitogenic Signaling Pathways... 96 5.3.3 Integrin and Growth Factor Receptor-Mediated Antiapoptotic Signaling Pathways... 97 5.3.4 The Anchorage Dependence... 99 5.3.5 Integrin and Growth Factor Receptor-Mediated Morphogenic Signaling Pathways... 100 5.3.6 Integrin-Mediated Mechanical Force-Induced Signaling... 105 5.4 Alterations in Integrin-Mediated Adhesion and Signaling in Transformed Cells... 108 5.4.1 Defective Adhesive Function... 108 5.4.2 Alterations in the Mitogenic Signal Transduction... 110 5.4.3 The Anchorage Independence... 111 References... 113 6 Cell Migration... 121 6.1 Factors Involved in Cell Migration... 121 6.1.1 Formation of Pseudopodia... 123 6.1.2 Polarization of Migrating Cells... 126 6.1.3 Contact Inhibition of Cell Migration... 127 6.1.4 Effect of Growth Factors... 128 6.1.5 Role of Focal Adhesions in Cell Migration... 130 6.2 Abnormalities of Cell Migration Machinery in Transformed Cells... 132 6.2.1 Pseudopodial Activity with Actin-Myosin Structure Deficiencies... 133 6.2.2 Cell-Matrix Adhesion Alterations... 134 6.2.3 Hypersensitivity to Mitogens-Motogens... 135 References... 137 7 Cell Responses to Chemical Heterogeneity of Substrata: Adhesive Islets or Paths... 145 References... 152 8 Topographic Cell Responses... 153 8.1 Cylindrical Substrata... 154 8.1.1 Normal Cell Responses... 154 8.1.2 Transformed Cell Responses... 158
Contents xv 8.2 Grooved Substrata... 161 8.2.1 Normal Cell Responses... 161 8.2.2 Transformed Cell Responses... 163 8.3 Discontinuous Substrata... 167 8.3.1 Lattices... 167 8.3.2 Multiple Vertvical Rods... 172 8.4 Effects of the Substratum Surface Topography on Cell Adhesion, Proliferation, and Synthetic Activities... 174 8.5 Mechanisms of Topographic Cell Responses... 177 8.5.1 Cylindrical Substrata... 178 8.5.2 Grooved Substrata... 179 8.5.3 Lattice Substrata... 180 References... 181 9 Intercellular Adhesive Interactions... 185 9.1 Cadherin-Mediated Intercellular Contacts: Adherens Junctions... 187 9.1.1 Structure of Adherence Junctions... 187 9.1.2 Dynamic Regulation of Adherens Junctions... 189 9.1.3 Contact Inhibition of Cell Proliferation... 197 9.2 Altered Regulation of Adherens Junctions Caused by Oncogenic Transformation... 200 9.2.1 Alterations in Cadherin Catenin Complex... 201 9.2.2 Loss of Contact Inhibition of Cell Proliferation... 204 References... 205 10 Conclusions... 213 References... 215 Index... 217
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Abbreviations ADAM (or MDC) proteins ADF ADP AMF APC Arp Arp2/3 Asef ATP ATPase Bad BAX Bcl2, Bcl-x BCL2, BCL-x BPAG1 (BP1230) CAM kinases camp Cas (p130 Cas) CDH1 c-erbb1/her1 CH-ILKBP c-met Cobl Cortactin CTNNB1 Metalloproteinase-like disintegrin- cystein rich proteins Actin-depolymerizing factor (cofilin) Adenosine diphosphate Autocrine motility factor Adenomatous polyposis coli protein Actin-related protein Actin-related protein complex Rac1-activating protein Adenosine triphosphate Adenosine triphosphatase Apoptogenic protein Apoptogenic tumor suppressor gene Antiapoptogenic proteins Antiapoptogenic proto-oncogenes Bullous pemphigoid antigen1 Calmodulin dependent protein kinases Cyclic adenosine monophosphate Focal adhesion protein Tumor suppressor gene encoding E-cadherin Proto-oncogene encoding EGF receptor Protein parvin Proto-oncogene encoding HGF/SF receptor Protein cordon-bleu Cortical actin-binding protein Proto-oncogene encoding b-catenin xvii
xviii Abbreviations DAG 1,2-diacylglycerol DAP kinase Death-associated protein kinase Dsh Disheveled protein EB1 (Microtubule plus-) end-binding protein1 EC1, EC5 Extracellular cadherin subdomains E-cadherin Epithelial cadherin ECM Extracellular matrix EGF Epidermal growth factor EMT Epithelial-mesenchymal transition Ena/VASP Enabled/vasodilator-stimulated phosphoprotein family ERM Ezrin, radixin and moezin protein family F-actin Filamentous actin FAK Focal adhesion tyrosine protein kinase FGF Fibroblast growth factor FH domain Formin homology domain FIP 200 Focal adhesion kinase family interaction protein of 200 kd FM Fluorescent microscopy formins Formin homology proteins G protein Guanine nucleotide-binding protein G-actin Globular actin GAP GTPase activating protein GDI GDP dissociation inhibitor GDP Guanosine diphosphate GEF Guanine nucleotide exchange factor GEF-H1 Rho guanine nucleotide exchange factor GF Growth factor GFAP Glial fibrillary acidic protein Girdin Girders of actin filaments protein GPCR G protein coupled receptor GSK3b Glycogen synthase kinase-3 b GTP Guanosine triphosphate GTPase Guanosine triphosphatase HGF/SF Hepatocyte growth factor, scatter factor IFs Intermediate filaments IGF-1 Insulin-like growth factor ILK Integrin-linked protein kinase INK4a Tumor suppressor gene IP3 Inositol 1,4,5-triphosphate IQGAP1 IQ motif-containing GTPase activating protein1 JMY Junction-mediated regulatory protein
Abbreviations LM Lmod MAP kinase (ERK) mdia MHC MLC MMP MRTF MSF MT1-MMP MTOC N-cadherin Necl NPF p140 Cap p27/kip1 p53 PAK P-cadherin PDGF PI PI3K PIK PINCH PIP PIP2 PIP3 PIPK PKA PKB (PKB/Akt) PKC PTEN Rab, Ras, Rho GTPases Rac, Rho, Cdc42 Rap1 R-cadherin RGD ROCK RTK Light microscopy Protein leiomodin Mitogen-activated protein kinase (extracellular signalregulated kinase) Formin homology protein Myosin heavy chain Myosin light chain Matrix metalloproteinase Myocardin-related transcription factor Migration stimulating factor Membrane type 1-matrix metalloproteinase Microtubule-organizing center (centrosome) Neural cadherin Nectin-like immunoglobulin-like adhesion molecule Nucleation-promoting factors xix Cas-associated protein Tumor suppressor gene Tumor suppressor gene, encoding p53 protein p21-activating protein kinase Placental cadherin Platelet-derived growth factor Phosphatidylinositol Phosphatidylinositol 3-kinase Phosphatidylinositol kinase Particularly interesting new cystein- histidine-rich protein Phosphoinositide Phosphatidylinositol biphosphate Phosphatidylinositol triphosphate Phosphatidylinositol phosphate kinase Protein kinase A, camp-dependent protein kinase Protein kinase B Protein kinase C Tumor suppressor gene, encoding phosphatase and tensin homolog (PTEN) protein Families of small GTPases Members of Rho family of small GTPases Member of Ras family of small GTPases Retinal cadherin Arginine-glycine-aspartic acid sequence Rho-associated kinase, Rho kinase Receptor tyrosine kinase
xx Scar(WAVE) SEM SMA Small G proteins SNAIL1 Tcf/Lef TGF-a TIAM1 TIMP +TIP VASP VE-cadherin VEGF WAF1 WASP WASP/WAVE (WASP/Scar) WAVE (Scar) WH2 WHAMM WIP Wnt Wnt1 glycoprotein XMAP215 Suppressor of camp receptor Scanning electron microscopy Alpha-smooth muscle actin Small GTPases, Ras superfamily GTPases Transcription repressor of CDH1 gene T-cell factor/lymphocyte- enhancer factor Transforming growth factor Rac-1 specific exchange factor Tissue inhibitor of metalloproteinase Microtubule plus-end tracking protein A member of Ena/VASP protein family Vascular endothelial cadherin Vascular endothelial growth factor Abbreviations p21 protein, a cell cycle inhibitor Wiscott-Aldrich syndrome protein family Wiscott-Aldrich syndrome protein (WASP) family that includes WASP family verprolin-homologous (WAVE) proteins WASP family verprolin-homologous proteins WASP-homolog 2 domain WASP homolog associated protein with actin, membranes, and microtubules WASP-interacting protein Protein family. The name Wnt is a combination of Wg ( wingless gene in Drosophila melanogaster) and Int (mouse oncogene) Protein encoded by WNT1 proto-oncogene Microtubule associated protein