Hexose transport in hybrids between malignant and normal cells

White, Marie; Bramwell, M.E.; Harris, Harriet A.

doi:10.1038/296588b0

Cited by 15 publications

(17 citation statements)

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“…The method was a modification of that employed by White et al (1981). Cells (10 ml-1), grown in 0.4ml volumes in the 16mm wells of plastic microtrays, were washed once with PBS pH 7.5 and incubated with 1 ml PBS containing 0.2 pCi 2-deoxy [3H] glucose (Amersham, 25 Ci mmol-1).…”

Section: Glucose Uptakementioning

confidence: 99%

Anti-tumour cytotoxin produced by human monocytes: Studies on its mode of action

Matthews¹

1983

Br J Cancer

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Summary Human monocytes can be induced to synthesize a cytotoxin which affects certain tumour cell lines. The interaction of monocyte cytotoxin with a susceptible cell line (L929) has been studied to obtain clues to the mode of action of the cytotoxin. The cytotoxin acts directly on the cells rather than on the culture medium and is cytotoxic at higher concentrations and cytostatic at lower concentrations. First signs of cell damage appear about 20 h after contact with the cytotoxin which must be present throughout this period. The cytotoxin probably acts on the cell surface and is more effective at 40°C than at 37°C. For a given amount of cytotoxin the effects are inversely proportional to the target cell concentration. Treatment of the cytotoxin with phenanthroline inhibits cytotoxicity while treatment of the target cells with actinomycin D, but not cycloheximide or puromycin, enhances cytotoxicity. After 24h cytotoxin treatment the target cells exhibit reduced respiration rate but enhanced glycolysis and glucose uptake suggesting mitochondrial dysfunction. A possible interpretation of these data is that the monocyte cytotoxin is a metalloenzyme which inactivates a cell surface receptor for a nutrient essential for mitochondrial function.A number of macrophage products with antitumour properties have been defined using in vitro assays. These products include arginase (Currie, 1978), cytolytic factor (Adams et al., 1980), tumour necrosis factor (Miinnel et al., 1980; Matthews, 1978, 198 la), human monocyte cytotoxin (Matthews, 1981b) and cytostatic factors CFI and CFII from human monocytes (Nissen-Meyer & Hammerstrqm, 1982). These factors may act separately or in concert (Adams et al., 1981;Nissen-Meyer and Hammerstr0m, 1982;Matthews, 1983).The human monocyte cytotoxin is apparently specific for certain tumour cell lines, it lacks species specificity and has a mol wt of -34,000 and slow electrophoretic mobility (Matthews, 1981b plastic adherence, were cultured overnight in Eagles minimum essential medium with 10% foetal calf serum (MEM/FCS) and 10 ugml-' endotoxin (Matthews, 1981b). A cytotoxin-enriched fraction was prepared from the monocyte supernatant by ion-exchange chromatography with CM-Sepharose (Matthews, 1983). Cytotoxin assay The mouse L929 tumour cell line was used as the target cells. Seventy-five pl amounts of target cell suspension (10 ml'1 in MEM/FCS) were pipetted into 96 well microtitre trays and incubated for at least 4h to allow the cells to adhere. Cytotoxin preparations were added in 75pl amounts, usually at 3 dilutions and with 3 or 4 replicates/dilution. After incubation at 37°C for 2-3 days the supernatant containing the dead cells was discarded and the adherent viable cells were fixed for 5min with 5% formaldehyde and stained with crystal violet. After drying, 100p1 33% acetic acid was added to each well to dissolve and evenly spread the dye. The amount of dye bound is proportional to the number of viable cells and was quantitated photometrically using a Titertek Multiskan photometer...

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Section: Glucose Uptakementioning

confidence: 99%

Anti-tumour cytotoxin produced by human monocytes: Studies on its mode of action

Matthews¹

1983

Br J Cancer

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“…These may be related to tissuespecific differences in glucose transport and may have different biochemical properties and physiological roles; for example, it has been reported that there is biochemical and functional heterogeneity in rat adipocyte glucose transporters (12) and that there are biochemical and functional differences in the hepatic and adipocyte glucose transporters (5). Changes in the expression of glucose transporter isozymes might account for differences in the Michaelis constants for hexose transport associated with certain tumors (28,29). We are currently cloning the cDNA for the chicken embryo fibroblast transporter message to gain a better understanding of the regulation of the chick transporter gene and to investigate this possibility.…”

Section: Methodsmentioning

confidence: 99%

Transformation by the src oncogene alters glucose transport into rat and chicken cells by different mechanisms.

White

Weber

1988

Mol. Cell. Biol.

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Transformation of both rat and chicken fibroblasts by the src oncogene leads to a four-to fivefold increase in the rate of glucose transport and in the level of the glucose transporter protein. We have previously shown that, with chicken embryo fibroblasts, transformation leads to a reduction in the rate of degradation of the transporter, with little or no increase in the rate of its biosynthesis. We now show that, with the rat-1 cell line, the opposite result was obtained. src-induced transformation led to an increase in transporter biosynthesis, with little effect on turnover. A src-induced increase in transporter mRNA entirely accounted for the increase in biosynthesis of the protein. By contrast, in chicken embryo fibroblasts, the level of transporter mRNA was low and was not induced to rise by src transformation. Thus, src induced an increase in the level of the glucose transport protein by fundamentally different mechanisms in chicken embryo fibroblasts and rat-1 cells. To test whether this difference was due to rat-1 cells being an immortalized cell line, we measured transporter mRNA levels in primary fibroblast cultures from rat embryos and in parallel cultures transformed by src. Transporter mRNA was inducible by src in these cells. Thus, the difference in mRNA inducibility between chicken and rat cells is not due to immortalization.Malignant transformation of chicken and rat embryo fibroblasts by Rous sarcoma virus (RSV) results in a marked increase in the rate of hexose transport across the plasma membrane (11,14,15,17,26,27). In cells transformed by RSV mutants in which the transforming protein pp60j-fsrc is temperature sensitive, the increase in transport is also temperature sensitive, indicating that the activity of pp6Ovsrc is necessary for both establishing and maintaining the transport alteration (14,15,17). It is generally accepted that the enhancement of transport is due to an increased number of hexose transporter molecules at the cell membrane, rather than a modification of existing carriers, for the following reasons. (i) Kinetic evidence indicates that the Vmax for transport is increased (7,14,15,26,27 biosynthesis showed that there was little or no difference between normal and transformed cells (22).In contrast, results obtained by others using rodent cells indicate that src transformation induces increased glucose transport by elevating the level of the mRNA for the transporter (8). To resolve this apparent discrepancy, we measured the effects of src transformation on the levels of transporter mRNA, transporter biosynthesis, and transporter degradation in chicken embryo fibroblasts, rat-i cells, and primary cultures of fibroblasts explanted from a rat embryo. We concluded that src increases the level of transporter by mechanisms that differ between rat and chicken cells. In the former, src elevates transporter mRNA levels and biosynthesis, whereas in the latter this is not the case and src acts to inhibit transporter turnover. 2) and stored at -20°C. Lysates were cleared by centrifugati...

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“…It is well established that stimulation of quiescent cells by mitogenic factors leads to an increased uptake of sugars and amino acids at an early stage in the sequence of events leading to an increased synthesis of DNA (for review, see Antoniades & Owen, 1982); cells transformed by oncogenic viruses (see Pasternak & Knox, 1977), and malignant as opposed to non-malignant cells (White et al, 1981), often show a similar increase. The infection of cells by cytolytic, nontransforming viruses such as vesicular stomatitis virus (VSV) or Semliki Forest virus (SFV) might be thought to have, if anything, an opposite effect on membrane function in that the synthesis of host DNA, RNA and protein becomes depressed (Pfefferkorn & Shapiro, 1974;Wagner, 1975), rather than elevated (but see Peluso et al, 1977).…”

Section: Introductionmentioning

confidence: 99%

Effect of Vesicular Stomatitis Virus and Semliki Forest Virus on Uptake of Nutrients and Intracellular Cation Concentration

Gray

Micklem

Brown

et al. 1983

Journal of General Virology

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Hexose transport in hybrids between malignant and normal cells

Cited by 15 publications

References 1 publication

Anti-tumour cytotoxin produced by human monocytes: Studies on its mode of action

Anti-tumour cytotoxin produced by human monocytes: Studies on its mode of action

Transformation by the src oncogene alters glucose transport into rat and chicken cells by different mechanisms.

Effect of Vesicular Stomatitis Virus and Semliki Forest Virus on Uptake of Nutrients and Intracellular Cation Concentration

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