A collection of readily reproducible experimental methods for studying the subversion of normal cell signaling that leads to cancer. Presented in step-by-step detail to ensure successful results, the methods include proven techniques for the investigation of apoptosis and cell death, complementary protocols for manipulating and/or monitoring oncogenic signals in cancer cells, and techniques for studying protein-protein interactions. A detailed protocol for capturing pure samples of malignant cells from frozen tissue specimens and two alternative techniques for analyzing their genomic DNA is also described.

Part I. Manipulation and Detection of Survival Signals. Functional Analysis of the Antimitogenic Activity of Tumor Suppressors, Erik S. Knudsen and Steven P. Angus. Rescue and Isolation of Rb-deficient Prostate Epithelium by Tissue Recombination, Simon W. Hayward, Yuzhuo Wang, and Mark L. Day. Signal Transduction Study Using Gene-Targeted Embryonic Stem Cells, Hideki Kawasome, Takashi Hamazaki, Tetsuo Minamino, and Naohiro Terada. The Use of the Yeast Two-Hybrid System to Measure Protein-Protein Interactions that Occur Following Oxidative Stress, Richard A. Franklin. Differential Screening of cDNA Libraries for Analysis of Gene Expression During Tumor Progression, Christopher W. Gregory. Mitogen-Activated Protein Kinase Signaling in Drug-Resistant Neuroblastoma Cells, Raymond R. Mattingly. TUNEL and Immunofluorescence Double-Labeling Assay for Apoptotic Cells with Specific Antigen(s), Stephanie M. Oberhaus. Part II. Manipulation and Detection of Oncogenic Signals. KinetworksTM Protein Kinase Multiblot Analysis, Steven Pelech, Catherine Sutter, and Hong Zhang. Protein Tyrosine Kinase and Phosphatase Expression Profiling in Human Cancers, Wen-Chang Lin. Association of Nonreceptor Tyrosine Kinase c-Yes with Tight Junction Protein Occludin by Coimmunoprecipitation Assay, Yan-Hua Chen and Qun Lu. Isolation of Novel Substrates Using a Tyrosine Kinase Overlay/In Situ Assay, Irwin H. Gelman. Manipulating Expression of Endogenous Oncogenic Proteins Using an Antisense Oligonucleotide Approach in Prostate Cancer Cells, Daqing Wu, Ginger G. Wescott, and David M. Terrian. Measurements of Phospholipases A2, C, and D (PLA2, PLC, and PLD): In Vitro Microassays, Analysis of Enzyme Isoforms, and Intact-Cell Assays, Julian Gomez-Cambronero, Joel Horwitz, and Ramadan I. Sha'afi. Detection of the Content and Activity of the Transcription Factor AP-1 in a Multistage Skin Carcinogenesis Model, Yunfeng Zhao and Daret K. St. Clair. Fibroblastic, Hematopoietic, and Hormone Responsive Epithelial Cell Lines and Culture Conditions for Elucidation of Signal Transduction and Drug Resistance Pathways by Gene Transfer, Linda S. Steelman, William L. Blalock, Xiao-Yang Wang, Phillip W. Moye, John T. Lee, John G. Shelton, Patrick M. Navolanic, Julianne M. Davis, Steven L. Knapp, Richard A. Franklin, Martyn K. White, and James A. McCubrey. Elucidation of Signal Transduction Pathways by Transfection of Cells with Modified Oncogenes, Linda S. Steelman, William L. Blalock, Xiao-Yang Wang, Phillip W. Moye, John T. Lee, John G. Shelton, Patrick M. Navolanic, Julianne M. Davis, Steven L. Knapp, Richard A. Franklin, Martyn K. White, and James A. McCubrey. Elucidation of Signal Transduction Pathways by Retroviral Infection of Cells with Modified Oncogenes, Linda S. Steelman, William L. Blalock, Xiao-Yang Wang, Phillip W. Moye, John T. Lee, John G. Shelton, Patrick M. Navolanic, Julianne M. Davis, Steven L. Knapp, Richard A. Franklin, Martyn K. White, and James A. McCubrey. Part III. Protein Interactions. Methods for the Study of Protein-Protein Interactions in Cancer Cell Biology, Daniel Price, Iha Park, and Hava Avraham. Production of Ligand-Specific Mutants Using a Yeast Two-Hybrid Mating Assay, André Nantel. Coimmunoprecipitation Assay for the Detection of Kinase-Substrate Interactions, Lucio Comai. Part IV. Genomic Rearrangements. Mutational Analysis of the Androgen Receptor Using Laser Capture Microdissection and Direct Sequencing, Sheila Greene, Patricia Stockton, Olga G. Kozyreva, Kris E. Gaston, Gordon P. Flake, and James L. Mohler. Clonality Analysis by T-Cell Receptor g PCR and High-Resolution Electrophoresis in the Diagnosis of Cutaneous T-Cell Lymphoma (CTCL), Ansgar Lukowsky. Index.