The contents of Colorectal Cancer: Methods and Protocols aim to instruct investigators in all the key genetic, cellular, and molecular biological methods of analyzing colorectal tumors. The focused techniques and assays are described in sufficient detail to allow researchers to start an experiment on colon tumors and proceed from beginning to end as if the expert in the field who has performed these studies were guiding them at the bench. Of note, most of the chapters in this volume are written by those scientists who p- neered these methods and assays in their respective fields. The chapters in Colorectal Cancer: Methods and Protocols describe state of the art methods to analyze colorectal tumors, ranging from gross mic- dissection of specimens to specific molecular analyses. Included are coverages of mutational assays, instability testing, immunohistochemical assays, chro- somal studies, and gene expression analyses. The goal of our volume is to facilitate the performance of colorectal tumor biological experiments by investigators at various levels of trainingfrom graduate students and postdoctoral fellows to principal investigators who desire to advance our understanding of colon cancer development.

Includes supplementary material: sn.pub/extras

Klappentext
Colorectal cancer is a common and highly lethal cancer once spread whose treatment is improving as we gain a better understanding of its development. In Colorectal Cancer: Methods and Protocols, Steven M. Powell, MD, has assembled a hands-on collection of cutting-edge molecular methods for studying this solid tumor. These readily reproducible techniques range from gross microdissection of specimens to specific molecular analyses. Here both the novice and experienced researcher will find productive chromosomal analyses, including comparative genomic hybridization and FISH assays, and revealing gene expression analyses, including immunohistochemical assays, Western blotting, and microarray assays. Also presented are mutational analyses of colon tumors and blood samples to determine whether specific patients have inherited a significant predisposition to colorectal cancer. Each method is described in step-by-step detail by an established expert to ensure robust experimental results, and all are accompanied by detailed notes on pitfalls to avoid.
Eminently practical, with each technique presented as if the expert in the field who had perfected it were guiding the investigator, Colorectal Cancer: Methods and Protocols provides cancer investigators with a gold standard collection of all the key genetic, cellular, and molecular biological methods for analyzing colon cancer today.

Inhalt
Microdissection of Histologic Sections.- Isolation of a Purified Epithelial Cell Population from Human Colon.- Xenografting Human Colon Cancers.- Comparative Genomic Hybridization Technique.- Fluorescence In Situ Hybridization.- Microsatellite Analysis to Assess Chromosome 18q Status in Colorectal Cancer.- The SURF Technique.- Microsatellite Instability Testing.- Immunohistochemical Analysis for hMLH1 and hMSH2 Expression in Colorectal Cancer.- Mutation Detection in Colorectal Cancers.- Mutation Detection inTGF-? Receptors.- Direct Analysis for Familial Adenomatous Polyposis Mutations.- I1307K Mutation Detection by Allele-Specific PCR in Familial Colorectal Cancer.- Microsatellite Analysis of the Insulin-Like Growth Factor II Receptor in Colorectal Carcinomas.- Molecular Detection of Smad2/Smad4 Alterations in Colorectal Tumors.- Direct Sequencing for Juvenile Polyposis Gene SMAD4/DPC4 Mutations.- 17 Direct Sequencing for Peutz-Jeghers Gene LKB1 (STK11) Mutations.- 18 Direct Sequencing for Cowden Syndrome Gene PTEN (MMAC1) Mutations.- Genetic Analysis Using Microarrays.- Cclooxygenase-2 (COX-2) Protein Expression by Western Blotting.- The Immunohistochemical Method.- Immunohistochemical Detection and Quantitation of Cell Surface Receptors for Prostanoids.- Adhesion Signaling Through Integrins.- Electrophoretic Mobility Shift Assay (EMSA).- Transfection Assays for Transformation of Model Colonic Cell Lines.- Assessment of Intestinal Stem Cell Survival Using the Microcolony Formation Assay.- Gene Transfer into the Colonic Mucosa.