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.
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