Ovarian Cancer: Methods and Protocols

ISBN-13: 9781489943965 / Angielski / Miękka / 2013 / 817 str.

John M. S. Bartlett

Opis

If there is one aspect of current cancer research that represents a major ch- lenge in both novice and experienced researchers, it is the rapid advance in our understanding of the disease. Researchers can be required to switch from analysis of gene expression to kinetics of protein activation, from genetic studies to the analysis of protein funtion. Cancers are highly complex disease systems and researchers aiming to understand the functioning of cancer systems require access to a wide range of laboratory techiques from a broad range of research disciplines. Increasingly, however, published methods are incomplete or refer back to a series of previous publications each containing only a small part of the complete pro- col. The aim of Ovarian Cancer: Methods and Protocols is to provide for ovarian cancer researchers in the first instance, a laboratory handbook that will facilitate research into cancer systems by providing a series of expert protocols, with proven efficacy, across a broad range of technical expertise. Thus, there are sections on tumor genetics and cellular signal transduction, as well as sections on apoptosis and RNA analysis. The value of Ovarian Cancer: Methods and Protocols to the ovarian cancer researcher will, I trust, be considerably enhanced by (1) the provision of a series of overviews relating to the biology, diagnosis, and treatment of this important neoplasm, and (2) the provision of a series of technical overviews introducing each part that provides an expert review of the applications and pitfalls of the various techniques included.

Spis treści

Part I. Introduction to Ovarian Cancer. The Epidemiology of Ovarian Cancer. Familial Ovarian Cancers. The Molecular Pathogenesis of Ovarian Cancer. Alterations in Oncogenes, Tumor Suppressor Genes, and Growth Factors Associated with Epithelial Ovarian Cancers. Pathological Assessment of Ovarian Cancer. Tumor Markers in Screeening for Ovarian Cancer. Primary Surgical Management of Ovarian Cancer. Recent Insights into Drug Resistance in Ovarian Cancer. Part II. Tumor Markers. Markers of Tumor Burden: An Overview. Bioactive Interleukin-6 Levels in Serum and Ascites as a Prognostic Factor in Patients with Epithelial Ovarian Cancer. ELISA Based Quantification of p105 (c-erb-B2, HER2/neu) in Serum of Ovarian Carcinoma. Enzyme Immunoassay of Urinary (-core Fragment of Human Chorionic Gonadotropin as a Tumor Marker for Ovarian Cancer. Part III. Model Systems. Ovarian Cancer Models: Technical Overview. Establishment of Ovarian Cancer Cell Lines. Subcloning of Ovarian Cancer Cell Lines. Culture and Characterization of Human Ovarian Surface Epithelium. MTT Growth Assays in Ovarian Cancer. In Vitro Invasion Assays. p53 Transfectants in Ovarian Cancer. Oestrogen Responsive Xenografts in Ovarian Cancer. The Use of Matrigel in the Establishment of Ovarian Carcinoma Cell Lines as Xenografts. Part IV. Cytogenetics. Cytogenetics of Ovarian Cancer: A Technical Overview. Interphase Cytogenetics in Frozen Ovarian Tumor Tissue. Interphase Cytogenetics in Paraffin Embedded Ovarian Tissue. Rapid Identification of Chromosomes Using Primed In Situ Labeling. Gene Amplification of c-ermB-2 detected by FISH. Chromosome Microdissection for Detection of Sub-Chromosomal Alterations by FISH. Quantitation of FISH Signals in Archival Tumors. Comparative Geneomic Hybridization in Paraffin Embedded Tissues. Part V. Molecular Genetic Imbalances in Ovarian Tumors. Molecular Genetics of Ovarian Cancer: A Technical Overview. Extraction of DNA from Microdissected Archival Tissues. Molecular Analysis of theUrokinase-Type Plasminogen Activator Gene. PCR Microsatellite Analysis of LOH in Ovarian Tumors. Molecular Genetic Analysis of Flow Sorted Ovarian and Microdissected Ovarian Tumor Cells: Improved Detection of Loss of Heterozygosity. SSCP and Sequence Analysis of p53 Mutation in Ovarian Tumors. Multiplex PCR (MPCR) Screening can Detect Small Intragenic p53 Deletion and Insertions. Transfer of Human-Chromosome 3 to an Ovarian Carcinoma Cell Line Identifies Regions Involved in Ovarian Cancer. Double Differential PCR and Competitive-Differential PCR for Gene Dosage Quantitation. One Dimensional Genome Scanning: Detection of Genomic Changes in Ovarian Carcinoma. Mapping of Tumor Suppressor Genes in Ovarian Cancer. Detection of the Replication Error Phenotype in Ovarian Cancer I-PCR Analysis of Microsatellite Instability. Immunostaining Human Paraffin Embedded Sections for Mismatch Repair Proteins. Quantitative PCR Detection of c-erbB-2 Gene Amplification. Part VI. mRNA Analysis. mRNA Anaysis: A Technical Overview. In Situ Hybridization Detection of TGRbeta mRNA. mRNA Detection by Insitu RT-PCR. RNase Protection Assay Analysis of mRNA for TGFb1-3 in Ovarian Tumors. RT-PCR Quantitation of HSP-60 mRNA Expression. The Effects of Butyrate and the Role of c-myc in N.1 Ovarian Carcinoma Cells. Differential Display. Part VII. Protein Expression. Measurement of Protein Expression: Technical Overview. Measurement of IGF-I Receptor Content in Tissues and Cell Lines aby Radioimmunoassay (RIA) and ELISA Techniques. Radioreceptor Measurement of ER/PR. Nm23 Protein Expression by Western Blotting in Patients with Epithelial Ovarian Carcinoma. Detection and Quantitation of Matrix Metalloproteases by Zymography. Simultaneous Flow Cytometric Detection of DNA and Cellular Protein Molecules. c-erbB-2 Immunohistochemistry in Paraffin Tumors. Quantitative Immunohistochemistry of Estrogen and Progesterone Receptor Positivity in Ovarian Tumors: A Rapid and Reliable Stereological Approach. Radioi

Blurb

Cancers are highly complex disease systems that require access to widely diverse laboratory techniques from a broad range of research disciplines, methods that are often beyond the knowledge of a single researcher. In Ovarian Cancer: Methods and Protocols, over one hundred well-respected scientists comprehensively review every aspect of the biology underlying ovarian cancer and present a detailed collection of easy-to-follow methods for illuminating this frequently fatal disease's basic mechanisms. Here today's investigators will find the broadest imaginable array of molecular biology techniques specially designed to determine not only tumor genetics, expression, and protein function, but also to elucidate the genetic mechanisms by which gene and immunotherapies may be perfected. The book's highly experienced authors provide help in selecting the correct model system for each experimental approach, and offer extensive technical advice to facilitate the planning of individual experiments and ensure that researchers are using the optimal techniques for their research programs. Although structured primarily for ovarian cancer researchers, and using examples from that field, the techniques can be immediately applied to research in many other tumor types.
Versatile and comprehensive, Ovarian Cancer Methods and Protocols constitutes a veritable treasury of well-proven expert methods that will help basic scientists and clinical researchers alike master the powerful technologies required for effective cancer research today.