Cancer Cell Biology: Methods and Protocols (Methods in Molecular Biology, 2508)

This volume provides detailed methods for the dissection of mechanisms underlying cancer
cell biology. This book begins with fundamental techniques and considerations for culturing
cancer cell lines and then more advanced techniques to study patient-derived tissue using
xenografts and cr yopreser vation of tumor cells. The book then describes methods to analyze the co-culture of breast cancer cells with adipocytes in two-dimensional and three-dimensional systems. Next are a series of chapters that allows the researcher to analyze cancer cell physiology including cell proliferation, protein secretion by ELISA, flow
cytometr y-based detection of protein levels by conventional fluorescence and mass spec-trometr y, and multi-parametric immunofluorescence analysis of tumor tissue. Also included are analysis of protein expression by wester n blot, analysis of sur face protein levels and recycling by biotinylation assay, and proteomics analysis by liquid chromatography mass spectrometry. This is followed by chapters describing a sophisticated analysis of cellular metabolism using the Seahorse assay, specific analysis of cancer cell mutations by CRISPR/Cas9, single cell RNA sequencing, along with a chapter on validation by reverse transcriptase quantitative PCR, and then bioinfor matics analysis of RNA-seq data. Lastly, chapters describing the isolation and characterization of extracellular vesicles follow.