hexacyanoferrate-iii and Cell-Transformation--Viral

It was found that the activities of prooxidant enzymes (NAD(P)H oxidases and NAD(P)H:cytochrome c reductases) in bovine leukemia virus-transformed calf and lamb embryo kidney fibroblasts (lines Mi-18 and FLK) were by 1.25-18 times higher when compared to corresponding nontransformed calf cells. The activity of DT-diaphorase was also increased by about one order of magnitude in transformed cells. The activities of antioxidant enzymes were almost unchanged (superoxide dismutase), decreased by 13% or 53% (catalase) or increased by 25% or 90% (glutathione reductase) in Mi-18 or FLK cells, respectively. These changes of enzyme activity increased the toxicity of simple redox-cycling quinones (duroquinone, naphthazarin) towards transformed cells, but did not affect the toxicity of daunorubicin. The latter was most probably related to the inhibition of plasma membrane NADH dehydrogenase.

Topics: Animals; Benzoquinones; Cattle; Cell Line, Transformed; Cell Survival; Cell Transformation, Viral; Embryo, Mammalian; Ferricyanides; Fibroblasts; Kidney; Leukemia Virus, Bovine; Multienzyme Complexes; NAD(P)H Dehydrogenase (Quinone); NADH Dehydrogenase; NADH, NADPH Oxidoreductases; NADPH Oxidases; NADPH-Ferrihemoprotein Reductase; Naphthoquinones; Oxidation-Reduction; Quinones; Sheep

Transformation of 3T3 cells by SV40 virus changes the properties of the transplasma membrane electron transport activity which can be assayed by reduction of external ferric salts. After 42 h of culture and before the growth rate is maximum, the transformed cells have a much slower rate of ferric reduction. The change in activity is expressed both by change in Km and Vmax for ferricyanide reduction. The change in activity is not based on surface charge effect or on tight coupling to proton release or on intracellular NADH concentration. With transformation by SV40 virus infection the expression of transferrin receptors increases, which correlates with greater diferric transferrin stimulation of the rate of ferric ammonium citrate reduction in transformed SV40-3T3 cells than in 3T3 cells.

Topics: 3T3 Cells; Animals; Cell Membrane; Cell Transformation, Viral; Electron Transport; Ferric Compounds; Ferricyanides; Kinetics; Mice; Oxidation-Reduction; Quaternary Ammonium Compounds; Simian virus 40; Transferrin

Changes have been found in the plasma membrane enzyme system which carries out transmembrane electron transport and associated proton transport in Simian virus 40 (SV40) temperature-sensitive A (tsA) mutant-transformed rat pineal cell line, RPN209-1. This cell line was temperature-sensitive for the maintenance of transformation. RPN209-1 cells expressed the transformed phenotype (rapid growth, high cell density, and cloning in soft agar) at the permissive temperature (33 degrees C) and the nontransformed phenotype (slower growth, lower saturation density, and lower cloning efficiency in soft agar) at the nonpermissive temperature (40 degrees C). The reduction of external ferricyanide, hexaammine ruthenium and diferric transferrin was used to measure the transmembrane redox activity. The transformed RPN209-1 cells expressed a lower transmembrane redox activity, which is more sensitive to the antitumor drug adriamycin, when compared to the cells with a nontransformed phenotype. The lower transmembrane redox activity is associated with a decrease in the affinity for ferricyanide and a change in Vmax of the enzyme. Since the transformed cells have 25% lower concentration of NADH, the decrease in Vmax may be partly based on substrate limitation. Ionic strength variation in the assay media shows that the change in activity with transformation is not based on change in cell-surface change. Treatment with neuraminidase, however, indicates that sialic acid is important for enzyme activity, consistent with previous proposals that the transmembrane enzyme is a glycoprotein. The proton extrusion associated with transplasma membrane electron transport is increased in transformed cells relative to the rate of ferricyanide reduction. A relation between proton pumping transplasma membrane electron transport and growth stimulation by external oxidants is discussed.

Topics: Animals; Cell Line; Cell Membrane; Cell Transformation, Viral; Electron Transport; Ferricyanides; Neuraminidase; Oxidation-Reduction; Pineal Gland; Rats; Simian virus 40; Temperature

Transplasma membrane electron transport activity by fetal rat liver cells (RLA209-15) infected with a temperature-sensitive strain of SV40 has been measured with cells grown at the restrictive temperature (40 degrees C) and permissive temperature (33 degrees C). The transformed cells grown at 33 degrees C had only one-half the rate of external ferricyanide reduction as the nontransformed cells held at 40 degrees C. Both the Km and Vmax for ferricyanide reduction were changed in the transformed state. The change in Vmax can be based on a decrease of NADH in the transformed cells. The change in rate with ferricyanide does not depend on change in surface charge. Reduction of external ferricyanide was accompanied by release of protons from the cells. The ratio of protons released to ferricyanide reduced was higher in the transformed cells than in the non-transformed cells. Since the transplasma membrane electron transport has been shown to stimulate cell growth under limiting serum, the changes in the plasma membrane electron transport and proton release in transformed cells may relate to modification of growth control.

Topics: Animals; Cell Division; Cell Membrane; Cell Transformation, Viral; Cells, Cultured; Electron Transport; Ferricyanides; Kinetics; Liver; NAD; Protons; Rats; Simian virus 40