Measuring the Impact of Microenvironmental Conditions on Mitochondrial Dehydrogenase Activity in Cultured Cells.
Mitochondria are powerhouses of a cell, producing much of the cellular ATP. However, mitochondrial enzymes also participate in many cellular biosynthetic processes. They are responsible for helping to maintain NAD(P)/H and redox balance, supplying metabolic intermediates for cell growth, and regulating several types of programed cell death. Several mitochondrial enzymes have even been shown to participate in the oncogenic process such as isocitrate dehydrogenase, succinate dehydrogenase, and fumarate hydratase. Recent advances have identified significant metabolic changes in the mitochondria that are regulated by malignant transformation and environmental stimuli. Understanding the biological activity and regulation of mitochondrial enzymes can provide insight into how they participate in the process of oncogenic transformation and work to sustain malignant growth. This chapter describes a technique to measure mitochondrial dehydrogenase activities that is faster and more cost effective which can also be scaled up for high throughput.
Center for Environmental and Systems Biochemistry, Mackey Cancer Center, University of KentucKY, Lexington, KY, 40536, USA.|Department of Radiation Oncology, Ohio State University Wexner School of Medicine and Comprehensive Cancer Center, Ohio Sate University, Columbus, OH, 43210, USA.
Ramon C
RC
Sun
Department of Radiation Oncology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Albert
A
Koong
Department of Radiation Oncology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Amato
A
Giaccia
Department of Radiation Oncology, Ohio State University Wexner School of Medicine and Comprehensive Cancer Center, Ohio Sate University, Columbus, OH, 43210, USA. Nicholas.denko@osumc.edu.
Nicholas C
NC
Denko
D045744:Cell Line, Tumor; D002478:Cells, Cultured; D060833:Cellular Microenvironment; D006801:Humans; D008928:Mitochondria / Q000201:enzymology*; D010088:Oxidoreductases / Q000378:metabolism*
D010088:Oxidoreductases
Colorimetric detection; Dehydrogenase; Enzymatic activity; Hypoglycemia; Hypoxia