Diverse molecular, cellular, and environmental events must all come together to allow the successful formation of secondary cancers, metastases. The second edition of Metastasis Research Protocols, brings together updated versions of the seminal technique that were presented in the first edition and also includes new techniques that have recently been shown to be important in illuminating the processes underlying this important area of biology. Volume 2 presents techniques applicable at the level of living cells and tissues, and presents methodologies applicable to cell behaviour in vitro, in animal models and in mathematical constructs. The aim is the study of the interaction between cancer cells and their host/environment. The focus throughout is on the tools that have been shown to be helpful in unravelling the processes important in cancer metastasis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Metastasis Research Protocols, Second Edition seeks to aid scientists in the further study of new methods in the area of metastasis research.

Diverse molecular, cellular, and environmental events must all come together to allow the successful formation of secondary cancers, metastases. The second edition of Metastasis Research Protocols, brings together the most up to date versions of the seminal techniques that were presented in the first edition and also includes new techniques that have recently been shown to be important in illuminating the processes underlying this important area of biology. Presented by top scientists, the collection includes a wide spectrum of articles encompassing important key methods and to introduce new methods which are making an impact in the area of metastasis research. Volume 1 includes key cellular and molecular techniques relevant to the exploration of cancer cells and tissues, the focus is on the tools that have been shown to be helpful in unravelling the molecular processes important in cancer metastasis. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Metastasis Research Protocols, Second Edition seeks to aid scientists in the further study of new methods in the area of metastasis research.

Diverse molecular, cellular, and environmental events must all come together to allow the successful formation of secondary cancers, metastases. The second edition of Metastasis Research Protocols, brings together updated versions of the seminal technique that were presented in the first edition and also includes new techniques that have recently been shown to be important in illuminating the processes underlying this important area of biology. Volume 2 presents techniques applicable at the level of living cells and tissues, and presents methodologies applicable to cell behaviour in vitro, in animal models and in mathematical constructs. The aim is the study of the interaction between cancer cells and their host/environment. The focus throughout is on the tools that have been shown to be helpful in unravelling the processes important in cancer metastasis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Metastasis Research Protocols, Second Edition seeks to aid scientists in the further study of new methods in the area of metastasis research.

Diverse molecular, cellular, and environmental events must all come together to allow the successful formation of secondary cancers, metastases. The second edition of Metastasis Research Protocols, brings together the most up to date versions of the seminal techniques that were presented in the first edition and also includes new techniques that have recently been shown to be important in illuminating the processes underlying this important area of biology. Presented by top scientists, the collection includes a wide spectrum of articles encompassing important key methods and to introduce new methods which are making an impact in the area of metastasis research. Volume 1 includes key cellular and molecular techniques relevant to the exploration of cancer cells and tissues, the focus is on the tools that have been shown to be helpful in unravelling the molecular processes important in cancer metastasis. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Metastasis Research Protocols, Second Edition seeks to aid scientists in the further study of new methods in the area of metastasis research.

The process of metastasis formation is hugely complex, as described in the introductory chapter of this book, and this complexity has led us to compile two volumes of methods, from a vastly divergent background that attempts to encompass the whole spectrum of cancer biology. This first volume, Metastasis Research Protocols: Analysis of Cells and Tissues, concentrates on analysis and mapping of molecules produced by cells and tissues and analysis of the molecular biology underlying their expression, whereas the second volume, Metastasis Research Protocols: Cell Behavior In Vitro and In Vivo,focuses sharply on the determination of cell behavior in vitro and in vivo. We have deliberately included chapters describing well-established and familiar te- niques (for example, SDS-PAGE and Western blotting [Chapter 11], and immunocytochemistry [Chapter 2]) in addition to the newer and more speci- ized approaches and specific examples of their application, because-although the methodology is readily available in the published literature and established in many laboratories-we wished these volumes to "stand alone" and to make accessible here the standard techniques that underpin much metastasis research for both the newcomer to the field and the seasoned researcher. Undoubtedly, owing to the complexity of the metastatic cascade and the wealth of research techniques involved in scientific approaches to its unraveling, and despite our best efforts to make these volumes as comprehensive as seems feasible, this is a tall order, and there will inevitably be omissions. For these we apologize.

In Volume I, Analysis of Cells and Tissues, we presented a range of protocols aimed at mapping and analyzing the expression of various molecules of pot- tial interest in metastasis research and for examining their production at the genetic level. In this second volume of metastasis research protocols, we move to the level of living cells and tissues and present methodologies applicable to examining metastatic behavior in vitro and in whole animal models. The methods described in the first section of this volume concentrate on the separation of cell lines with high and low metastatic potential, including the genetic modification of cell lines. The assay systems to test defined aspects of the metastatic cascade are then described in Part II and include cell migration assays, assays for matrix degrading enzymes, basement membrane degrading assays, adhesion assays, and assays of angiogenesis. The role of the specific elements of the metastatic cascade assayed in each of these systems in turn must of course be put into perspective relative to their roles in entire living organisms.

A collection of both well-established and cutting-edge methods for investigating breast cancer biology not only in the laboratory, but also in clinical settings. These readily reproducible techniques solve a variety of problems, ranging from how to collect, store, and prepare human breast tumor samples for analysis, to analyzing cells in vivo and in vitro. Additional chapters address the technology of handling biopsies, new methods for analyzing genes and gene expression, markers of clinical outcome and progress, analysis of tumor-derived proteins and antigens, validating targets, and investigating the biology of newly discovered genes.