ToxSci Advance Access published online on March 14, 2007
Toxicological Sciences, doi:10.1093/toxsci/kfm052
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Circumventing the Crabtree Effect: Replacing Media Glucose with Galactose Increases Susceptibility of HepG2 Cells to Mitochondrial Toxicants
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* Pfizer DSRD, 10646 Science Center Drive, San Diego, CA 92121
Luxcel Biosciences Ltd., BioInnovation Centre, BioTransfer Unit, UCC, Cork Ireland
2 corresponding author: Yvonne Will, Ph.D., Pfizer DSRD, 10646 Science Center Drive, San Diego, CA 92121, Telephone: 858-622-7544, FAX: 858-678-8290, Email: Yvonne.Will{at}pfizer.com
Received January 12, 2007; revision received March 8, 2007; accepted March 8, 2007
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Abstract |
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Many highly proliferative cells generate almost all ATP via glycolysis despite abundant O2 and a normal complement of fully-functional mitochondria, a circumstance known as the Crabtree Effect. Such anaerobically poised cells are resistant to xenobiotics that impair mitochondrial function, such as the inhibitors rotenone, antimycin, oligomycin and compounds like FCCP, that uncouple the respiratory electron transfer system from phosphorylation. These cells are also resistant to the toxicity of many drugs whose deleterious side effect profiles are either caused, or exacerbated, by impairment of mitochondrial function. Drug-induced mitochondrial toxicity is shown by members of important drug classes, including the thiazolidinediones, statins, fibrates, antivirals, antibiotics, and anticancer agents. To increase detection of drug-induced mitochondrial effects in a preclinical cell-based assay, HepG2 cells were forced to rely on mitochondrial oxidative phosphorylation rather than glycolysis by substituting galactose for glucose in the growth media. Oxygen consumption doubles in galactose-grown HepG2 cells, and their susceptibility to canonical mitochondrial toxicants correspondingly increases. Similarly, toxicity of several drugs with known mitochondrial liabilities is more readily apparent in aerobically-poised HepG2 cells compared to glucose-grown cells. Some drugs were equally toxic to both glucose- and galactose-grown cells, suggesting that mitochondrial impairment is likely secondary to other cytotoxic mechanisms.
Key Words: Mitochondria; Drug Safety Testing; HepG2 cells; respiration; OXPHOS.
1 These authors contributed equally to the work presented
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