Metastasis Research Protocols

ISBN: 978-0-89603-610-9
Verlag: Springer, Berlin

Buch, Englisch, Band 57, 344 Seiten, Format (B × H): 152 mm x 229 mm, Gewicht: 680 g

Reihe: Methods in Molecular Medicine

Metastasis Research Protocols describes in detail all the methods needed to investigate why and how metastasis occurs. Volume I presents a comprehensive collection of established and leading-edge techniques for analyzing the expression of key molecules and for examining their production at the genetic level. The work focuses on the analysis and mapping of molecules produced by cells and tissues, and on the molecular biology underlying their expression. The traditional methods range from the histopathological and the immunocytochemical to SDS-PAGE, Western blotting, and enzyme zymography. Newer and more specialized techniques for analyzing the genetic aspects of metastasis include in situ hybridization to localize mRNAs, FISH, CGH, methylation analysis of CpG islands, RT-PCR, and differential display. Volume II presents a comprehensive collection of established and leading-edge techniques for examining metastatic behavior in vitro and whole animal models. Methods are provided for the separation of cell lines with high and low metastatic potential, along with assay systems to test defined aspects of the metastatic cascade. These systems include cell migration assays, assays for matrix degrading enzymes, basement membrane degrading assays, adhesion assays, and assays of angiogenesis. The detailed descriptions of animal models of metastasis cover the use of immunodeficient animals, syngenic and transgenic models, and orthotopic models of metastasis. Although ninety percent of fatal cancer cases involve the spread of a primary tumor, the formation of metastases is still a poorly understood, complex process and a significant problem in the treatment of cancer patients. In Metastasis Research Protocols, leading international investigators describe the key methods needed to investigate why and how metastasis occurs. Volume I of this two-volume set, Analysis of Cells and Tissues, presents a comprehensive collection of established and leading-edge techniques for analyzing the expression of key molecules and for examining their production at the genetic level. The work focuses on the analysis and mapping of molecules produced by cells and tissues, and on the molecular biology underlying their expression. The traditional methods range from the histopathological and the immunocytochemical to SDS-PAGE, Western blotting, and enzyme zymography. Newer and more specialized techniques for analyzing the genetic aspects of metastasis include in situ hybridization to localize mRNAs, FISH, CGH, methylation analysis of CpG islands, RT-PCR, and differential display. The second volume of this set, Analysis of Cell Behavior In Vitro and In Vivo, moves to the level of living cells and tissues to present methodologies applicable to examining metastatic behavior in vitro and in the whole animal.
Comprehensive and authoritative, the two volumes of Metastasis Research Protocols constitute a gold-standard collection of readily reproducible methods for understanding the metastatic cascade-first at the cellular and molecular levels, then at the level of the whole organism-responsible for the spread of cancer and for developing novel strategies to combat its spread.

Autoren/Hrsg.

Brooks, Susan A.

Schumacher, Udo

Inhaltsverzeichnis

Part I. Detection Methods for Cellular Markers of Metastatic Potential
Histopathological Assessment of Metastasis
Derek E. Roskell and Ian D. Buley
Basic Immunocytochemistry for Light Microscopy
Susan A. Brooks
Multiple Labeling Techniques for Fluorescence Microscopy
Amanda J. Atherton and Catherine Clarke
Lectin Histochemistry to Detect Altered Glycosylation in Cells and Tissues
Susan A. Brooks and Debbie M. S. Hall
Immunocytochemical Detection and Characterization of Individual Micrometastatic Tumor Cells
Stephan Braun and Klaus Pantel
Understanding Metastasis Through Integrin Expression
Lawrence John and Massimo Pignatelli
Surrogate Markers of Angiogenesis and Metastasis
Girolamo Ranieri and Giampietro Gasparini
Measurement of Microvessel Density in Primary Tumors
Helen E. Turner and Adrian L. Harris
Assessment of Cellular Proliferation by Calculation of Mitotic Index and by Immunohistochemistry
Cheryl E. Gillett
Flow Cytometric Measurement of Cell Proliferation
Richard S. Camplejohn
SDS-PAGE and Western Blotting Techniques
Christine Blancher and Adam Jones
Assessment of Gelatinases (MMP-2 and MMP-9) by Gelatin Zymography
Marta Toth and Rafael Fridman
Part II. Genetic Aspects of Metastasis
In Situ Hybridization to Localize mRNAs
Richard Poulsom
Fluorescence In Situ Hybridization
Hakan Goker and Janet Shipley
Comparative Genomic Hybridization
Rebecca Roylance
RNA Purification and Quantification Methods
Adam Jones, Chisato Fujiyama, and Kenneth Smith
Northern and Southern Blotting and the Polymerase Chain Reaction to Detect Gene Expression
Thelma Tennant, Marina Chekmareva, and Carrie W. Rinker-Schaeffer
Methylation Analysis of CpG Islands
Richard Lilischkis, Hermann Kneitz, and Hans Kreipe
Detection of Metastatic Tumor Cells in Blood by Reverse Transcriptase-Polymerase Chain Reaction
Katja Haack, Krishna Soondrum, and Michael Neumaier
Differential Display
Ruey Ho Kao and Giulio Francia
Part III. Mathematical Modeling of Metastases
Predictive Mathematical Modeling in Metastasis
Jonathan A. Sherratt