Investigation of the carbohydrate metabolism of normal and neoplastic hepatocytes using 2,6-dichlorophenolindophenol as a probe for NAD(P)H production measured by voltammetry.
An electrochemical technique is described for measurement of intracellular NAD(P)H production. This technique involves an auxiliary redox system taken up by the cells which is then measured voltammetrically after reduction by NAD(P)H. The redox system used was 2, 6-dichlorophenolindophenol (DCPIP). It was shown to undergo a quasi-reversible two-electron transfer at the rotating gold disc electrode serving as an indicator electrode. The anodic wave of the reduced form of DCPIP was taken to indicate the amount of NAD(P)H produced by metabolic processes with glucose as substrate for a given number of cells. The following types of cell were investigated in suspension: Morris hepatoma 3924, a hepatocyte-derived cell line, and normal hepatocytes. Marked differences between normal and transformed cells were found under aerobic compared to anaerobic conditions. These were explained in terms of alterations in carbohydrate metabolism, e.g., Pasteur effect occurring on cell transformation.
D015086:2,6-Dichloroindophenol / Q000378:metabolism*; D000818:Animals; D050260:Carbohydrate Metabolism; D002460:Cell Line; D004396:Coloring Agents; D004563:Electrochemistry; D005947:Glucose / Q000378:metabolism; D007215:Indophenol / Q000031:analogs & derivatives*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008297:Male; D009243:NAD / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000494:pharmacology; D011199:Potentiometry; D051381:Rats; D011919:Rats, Inbred Strains
D004396:Coloring Agents; D009243:NAD; D007215:Indophenol; D009249:NADP; D015086:2,6-Dichloroindophenol; D005947:Glucose; D010100:Oxygen