Effect of glucose and deoxyglucose on the redistribution of calcium in ehrlich ascites tumour and Zajdela hepatoma cells and its consequences for mitochondrial energetics. Further arguments for the role of Ca(2+) in the mechanism of the crabtree effect.
The distribution of Ca(2+) in intact cells was monitored with fluorescent probes: fura-2 for cytosolic [Ca(2+)] and rhod-2 for mitochondrial [Ca(2+)]. It was found that in neoplastic cells, such as Ehrlich ascites tumour and Zajdela hepatoma, but not in non-malignant cells, such as fibroblasts, glucose and deoxyglucose elicited release of Ca(2+) from endoplasmic reticulum stores and an increase in Ca(2+) concentration in the cytosol. Parallel to this, a decrease in the rate of Ca(2+) extrusion from the cell and an enhanced uptake of Ca(2+) by mitochondria were observed. The increase in mitochondrial [Ca(2+)] was accompanied by an increase in the mitochondrial membrane potential and the reduction state of nicotinamide nucleotides. F(1)F(o)-ATPase in submitochondrial particles of Zajdela hepatoma was strongly inhibited in the presence of micromolar Ca(2+) concentrations, whereas this activity in submitochondrial particles from rat liver appeared to be less sensitive to Ca(2+). Indications of glycosylation of Ehrlich ascites tumour cell proteins were also obtained. These data strengthen the proposal [Bogucka, K., Teplova, V.V., Wojtczak, L. and Evtodienko, Y. V. (1995) Biochim. Biophys. Acta 1228, 261-266] that the Crabtree effect is produced by mobilization of cell calcium, which is subsequently taken up by mitochondria and inhibits F(1)F(o)-ATP synthase.
Nencki Institute of Experimental Biology, Warsaw, Poland Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Russian Federation. LWAC@nencki.gov.pl
L
L
Wojtczak
V V
VV
Teplova
K
K
Bogucka
A
A
Czyz
A
A
Makowska
M R
MR
Wieckowski
J
J
Duszyński
Y V
YV
Evtodienko
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology / Q000502:physiology*; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism*; D003847:Deoxyglucose / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013053:Spectrophotometry; D019284:Thapsigargin / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
D004791:Enzyme Inhibitors; D019284:Thapsigargin; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D005947:Glucose; D002118:Calcium