Flow-cytometric determination of dehydrogenase activities in primary human gastrointestinal tumor cell lines

Abstract

The activities of 6 dehydrogenases, lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (ICDH), glycerol-3-phosphate dehydrogenase (GDH), succinate dehydrogenase (SDH) and glutamate dehydrogenase (GLDH), determined by means of flow cytometry in 13 primary human gastrointestinal tumour cell lines, including 10 esophageal carcinomas, one gastric cancer, and 2 pancreatic cancers. Two-parametric measurements of specific dehydrogenase activities in single cells were performed with DAPI as fluorochrome for the nuclear DNA and with the fluorescent redox system of 5-cyano-2,3-ditolyl-tetrazolium chloride (CTC) which forms brilliant red formazan crystals upon reduction by cellular redox enzymes. Furthermore, with the aid of the calibration procedure reported previously [18] the enzyme activities were expressed as biochemical units. This application of tetrazolium salt technique for demonstrating dehydrogenase activities in human tumour cells by flow cytometry offers an alternative tool to characterize malignant tumors.