N,N-Dimethylformamide modulates acid extrusion from murine hepatoma cells

N,N-Dimethylformamide (DMF) affects cellular differentiation, causes hepatotoxicity and gastric irritation, and may be carcinogenic. Since these processes involve changes in cellular pH homeostasis, we investigated the effects of DMF on H+ extrusion and cytosolic pH (pHi) of mouse hepatoma cells (Hepa 1C1C7). Extracellular pH was monitored using a silicon-based sensor system (Cytosensor microphysiometer) and pHi was monitored by fluorescence spectrophotometry. Superfusion of cells with DMF (0.25 to 0.5 M) suppressed the extracellular acidification rate (ECAR) below baseline. Following washout of DMF there was a rapid, concentration-dependent, prolonged overshoot of ECAR above baseline rates. Removal of extracellular Na+ or superfusion with amiloride abolished the overshoot in acidification rate, indicating involvement of Na+/H+ exchange. The overshoot was dependent on extracellular glucose, suggesting that it arises from an increase in metabolic acid production. Fluorescence measurements showed that DMF did not change pHi. Furthermore, DMF did not alter the rate of pHi recovery of cells acid loaded using nigericin, indicating that DMF does not directly alter Na+/H+ exchange activity in these cells. In summary, these data suggest that suppression of acidification rate by DMF is likely due to decreased metabolic acid production. Washout of DMF is then accompanied by increased glucose metabolism and H+ efflux via Na+/H+ exchange. It is possible that alterations in H+ production and transport contribute to the hepatotoxicity of DMF and its effects on cellular differentiation.