Full Description
This volume includes comprehensive methods and protocols on autophagic pathways in cancer disorders. Chapters cover basic principles of autophagy methodology in cancer cells, translational approaches, systematic computational analyses of TCGA (The Cancer Genome Atlas) cohorts, CCLE (The Cancer Cell Line Encyclopedia) analysis, CRISPR genetics, metabolomics, biochemistry, and immunohistochemistry methodologies. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Autophagy and Cancer: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field.
Contents
A balance between autophagy and other cell death modalities in cancer.- Isolation of Autophagy competent Lysosomes from Cancer cells by Differential Large-scale Multi-layered Density Gradient Centrifugations.- CRISPR-Cas9 Gene Editing to create Isoform specific LAMP-2A Knockout in Human Cancer Cells.- Modified LC3 dot quantification method.- A quantitative flow cytometry-based method for autophagy detection across the cell cycle.- A dual HiBiT-GFP-LC3 lentiviral reporter for autophagy flux assessment.- Measuring autophagic cargo flux with Keima-based probes.- Detection of nuclear biomarkers for chromosomal instability.- Analysis of autophagic vesicles in mitotic cells.- Approaches and protocols to analyze autophagy and its role in death of apoptosis-resistant senescent tumor cells.- Concepts and design of introducing synthetic microRNAs into mammalian cells.- Studying autophagy using a TMT-based quantitative proteomics approach.- Immunofluorescence-based measurement of autophagosome formation during mitophagy.- Induction and detection of mitophagy.- Analysis of pancreatic acinar protein solubility in autophagy-deficient mice.- Studying autophagy in vivo in the mammary gland and in xenograft samples.- A method to Identify Potential Prognostic Markers Across Distinct Tumor Types.- A method for Co-Expression Analysis.- Methodological approaches for assessing metabolomic changes in glioblastomas -MS.- Extraction of Metabolites from Cancer Cells.- Measuring glucose consumption and gluconeogenesis in 3D human tissue cultures with nanoliter input volumes.
