Full Description
Volume 542 of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This new volume covers research methods providing a theoretical overview on metabolic alterations of cancer cells and a series of protocols that can be employed to study oncometabolism, in vitro, ex vivo and in vivo. Malignant cells exhibit metabolic changes when compared to their normal counterparts, owing to both genetic and epigenetic alterations. Although such a metabolic rewiring has recently been indicated as "yet another" general hallmark of cancer, accumulating evidence suggests that the metabolic alterations of each neoplasm rather represent a molecular signature that intimately accompanies, and hence cannot be severed from, all facets of malignant transformation.
Contents
1. The metabolic alterations of cancer cells
2. Autophagy and cancer metabolism
3. Regulation of cancer metabolism by oncogenes and tumor suppressors
4. Crosstalk between cell death regulation and metabolism
5. Techniques to monitor glycolysis
6. Measurement of enolase activity in cell lysates
7. Extracellular flux analysis to monitor glycolytic rates and mitochondrial oxygen consumption
8. Conventional techniques to monitor mitochondrial oxygen consumption
9. Combined high-resolution respirometry and fluorometry. Validation of safranin for determination of mitochondrial membrane potential.
10. Kinetic analysis of local oxygenation and respiratory responses of mammalian cells using intracellular oxygen-sensitive probes and time-resolved fluorescence plate reader
11. 13C isotope-assisted methods for quantifying glutamine metabolism in cancer cells
12. Measurement of fatty acid oxidation rates in animal tissues and cell lines
13. Methods to assess steroid hormone induced lipid accumulation in cancer cells
14.Use of chemical probes to monitor mitochondrial ROS production by fluorescence dyes for flow cytometry and spectrofluorometry
15. Rapid methods to monitor mitochondrial and cellular ROS production by fluorescence microscopy and fluorometry
16. Genetically encoded redox sensors in biomedical science
17. Real time measurement of metabolic states in living cells using genetically encoded NADH sensors
18. Use of genetically encoded sensors to monitor cytosolic ATP/ADP ratio in living cells
19.Methods to monitor and compare mitochondrial and glycolytic ATP production in cells
20. Measurement of ADP-ATP exchange in relation to membrane potential and oxygen consumption in mitochondria with improved calibration
21. Analysis of hypoxia-induced metabolic reprogramming
