Phorbol ester stimulation of Na influx and Na-K pump activity in swiss 3T3 cells

Dicker, Phillip; Rozengurt, Enrique

doi:10.1016/s0006-291x(81)80115-5

Cited by 105 publications

(40 citation statements)

References 15 publications

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“…Of particular importance to this study is the observation that PMA is mitogenic in quiescent cultures of this cell line (12), as it is for other 3T3 cell lines (8, 9). In contrast to the earlier work of others (8,9) showing an increased rate of 'Rb+ uptake caused by PMA, we found in these cells that inhibition of 86Rb+ uptake was an early event after PMA treatment. The transport system inhibited by PMA in these cells is insensitive to ouabain and sensitive to the diuretic furosemide, and it apparently catalyzes the cotransport of Na+, K+, and Cl-.…”

contrasting

confidence: 99%

“…Moroney et al (8) reported that an early response of PMA-treated Swiss 3T3 cells was a stimulation of ouabain-sensitive 86Rb+ uptake (i.e., Na+/K+ pump-mediated). Dicker and Rozengurt (9) later confirmed this finding and suggested that the stimulation of pump activity by PMA was caused by an increased Na+ influx, although the mechanism of this effect was not specified.…”

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confidence: 98%

“…This cell line is well suited for these studies because previous work has detailed numerous responses to PMA, including stimulation of sugar transport activity (10), increased rate of glycolysis (11), and inhibition of cell differentiation (12). Of particular importance to this study is the observation that PMA is mitogenic in quiescent cultures of this cell line (12), as it is for other 3T3 cell lines (8,9). In contrast to the earlier work of others (8,9) showing an increased rate of 'Rb+ uptake caused by PMA, we found in these cells that inhibition of 86Rb+ uptake was an early event after PMA treatment.…”

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confidence: 99%

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Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

O’Brien¹,

Krzeminski²

1983

Proc. Natl. Acad. Sci. U.S.A.

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The tumor promoter phorbol 12-myristate 13-acetate (PMA) rapidly decreased the rate of "Rb+ uptake into BALB/c 3T3 preadipose cells. The component of total 86Rb+ influx affected by PMA is insensitive to ouabain but sensitive to the diuretic furosemide. Experiments designed to investigate the characteristics of the K+ transport system sensitive to PMA revealed that: (i) 86Rb+ uptake is highly dependent on external Na+, (ii) "Rb+ uptake is highly dependent on external ClP, (iii) 22Na+ uptake is dependent on external K+, and (iv) a major component of 86Rb+ efflux that is sensitive to PMA and furosemide is not dependent on extracellular KV. These features strongly implicate a Na+K+/Cl-cotransport system as the target of PMA and furosemide in these experiments. PMA caused a net intracellular accumulation of K+ within 15 min in these cells, presumably via its inhibitory effect on furosemide-sensitive K+ transport. Within 30 min after PMA treatment, the mean cell volume was significantly reduced in treated compared to control cells, with a maximum decrease of 21% attained at 4 hr after PMA. The significance of these findings for biologic changes induced by PMA is discussed.In addition to their role in multistage carcinogenesis, the phorbol ester tumor promoters, exemplified by phorbol 12-myristate 13-acetate (PMA), cause a number of biologic and functional changes in diverse cell types in culture (for reviews, see refs. 1 and 2). To date, however, no experimental results to explain any of the actions of tumor promoters in (11), and inhibition of cell differentiation (12). Of particular importance to this study is the observation that PMA is mitogenic in quiescent cultures of this cell line (12), as it is for other 3T3 cell lines (8, 9). In contrast to the earlier work of others (8,9) showing an increased rate of 'Rb+ uptake caused by PMA, we found in these cells that inhibition of 86Rb+ uptake was an early event after PMA treatment. The transport system inhibited by PMA in these cells is insensitive to ouabain and sensitive to the diuretic furosemide, and it apparently catalyzes the cotransport of Na+, K+, and Cl-.MATERIALS AND METHODS Cells. BALB/c 3T3 cells (clone A3IT) were obtained from Leila Diamond (Wistar Institute) and cultured as described (12). The medium used was Eagle Auto-Pow minimal essential medium (Flow Laboratories) supplemented with 10% fetal bovine serum. For flux and ion content experiments, cells were plated from stocks into 35-or 60-mm plastic Petri dishes and used when growing logarithmically or just after reaching confluence. Routinely, cultures were refed with fresh medium every 2-3 days and always on the day prior to experiments.Flux Experiments. The uptake of 'Rb+ was measured in cells maintained in complete culture medium at 37°C in a humidified atmosphere of 95% air/5% CO2. Cells in 35-mm dishes were washed with warm phosphate-buffered saline (unless otherwise stated) and uptake was started by the addition of 2 ml of fresh medium containing 10% fetal bovine serum (K+, 5.4 mM)...

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Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

O’Brien¹,

Krzeminski²

1983

Proc. Natl. Acad. Sci. U.S.A.

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“…Furthermore, it has been reported that inhibition of neuronal NE reuptake is not responsible for the phorbol ester-potentiated NE release (30,31). PKC activation causes blockade of K+ currents (23) or facilitation of Na+ (32) and Ca 2+ (33) currents, each of which results in an enhancement of NE re lease. The present study with TEA, a K+ channel blocker, reveals that blockade of K+ channels is not responsible for the 40 mM KCI-evoked tritium overflow mechanisms.…”

Section: Discussionmentioning

confidence: 99%

Presynaptic Sites of Isolated Canine Saphenous Veins Are More Sensitive to Protein Kinase C than Postsynaptic Ones.

1991

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“…The pleiotypic responses include increased transport of ions and hexoses (18,19,29,39,41), increased activity of rate-limiting enzymes in anabolic pathways (e.g., ornithine decarboxylase) (33,34), and increased phosphorylation of a number of cellular proteins (28,36,39). Of particular relevance to the work reported in this communication are the findings that both EGF and TPA cause increased serine/threonine phosphorylation of the EGF receptor (11,17,20,23), that TPA stimulates phosphorylation of an 80-kDa soluble protein (36), and that all the mitogens (including TPA) cause tyrosyl phosphorylation of a 42-kDa cellular protein (4,16,21,31).…”

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confidence: 99%

Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells.

Bishop¹,

Martinez²,

Weber³

et al. 1985

Mol. Cell. Biol.

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The 3T3-TNR9 cell line is a variant of Swiss 3T3 cells which does not respond mitogenically to tumor promoters, but does respond mitogenically to epidermal growth factor, fibroblast growth factor, and serum. To elucidate differences between tumor promoters and polypeptide mitogens in the pathway(s) of mitogenesis which might be responsible for the nonresponsiveness of the 3T3-TNR9 cells, we have examined in these cells the early protein phosphorylation events known to be associated with mitogenesis in the parental 3T3 cells. We find that the 3T3-TNR9 cells display levels of tetradecanoyl phorbol acetate binding and of a calcium- and phospholipid-dependent protein kinase activity which are at least the equal of those seen in the parental 3T3 cells, implicating some postreceptor event in the nonmitogenic phenotype. In addition, we find that phosphorylation of the epidermal growth factor receptor and of 80-kDa and 22-kDa proteins, as well as the tyrosine phosphorylation of a 42-kDa protein, all proceed normally in the nonmitogenic variant, even though these phosphorylations must depend on the activation of different kinases. Thus, all these early phosphorylation reactions are intact in the 3T3-TNR9 cells. Although these phosphorylations may be necessary, they clearly are insufficient to trigger mitogenesis.

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Phorbol ester stimulation of Na influx and Na-K pump activity in swiss 3T3 cells

Cited by 105 publications

References 15 publications

Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

Phorbol ester inhibits furosemide-sensitive potassium transport in BALB/c 3T3 preadipose cells.

Presynaptic Sites of Isolated Canine Saphenous Veins Are More Sensitive to Protein Kinase C than Postsynaptic Ones.

Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells.

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