Full Description
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 training—from graduate students and postdoctoral fellows to principal investigators who desire to advance our understanding of colon cancer development.
Contents
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.
