Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Jul;80(14):4334–4338. doi: 10.1073/pnas.80.14.4334

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

T G O'Brien, K Krzeminski
PMCID: PMC384032  PMID: 6576341

Abstract

The tumor promoter phorbol 12-myristate 13-acetate (PMA) rapidly decreased the rate of 86Rb+ 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) 86Rb+ uptake is highly dependent on external Cl-, (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 K+. 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.

Full text

PDF
4334

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blumberg P. M. In vitro studies on the mode of action of the phorbol esters, potent tumor promoters, part 2. Crit Rev Toxicol. 1981 Jun;8(3):199–234. doi: 10.3109/10408448109109658. [DOI] [PubMed] [Google Scholar]
  2. Boynton A. L., Whitfield J. F., Isaacs R. J. Calcium-dependent stimulation of BALB/c 3T3 mouse cell DNA synthesis by a tumor-promoting phorbol ester (PMA). J Cell Physiol. 1976 Jan;87(1):25–32. doi: 10.1002/jcp.1040870105. [DOI] [PubMed] [Google Scholar]
  3. Boynton A. L., Whitfield J. F. Stimulation of DNA synthesis in calcium-deprived T51B liver cells by the tumor promoters phenobarbital, saccharin, and 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1980 Dec;40(12):4541–4545. [PubMed] [Google Scholar]
  4. Diamond L., O'Brien S., Donaldson C., Shimizu Y. Growth stimulation of human diploid fibro-blasts by the tumor promoter, 12-0-tetradecanoly-phorbol-13-acetate. Int J Cancer. 1974 May 15;13(5):721–730. doi: 10.1002/ijc.2910130516. [DOI] [PubMed] [Google Scholar]
  5. Diamond L., O'Brien T. G., Baird W. M. Tumor promoters and the mechanism of tumor promotion. Adv Cancer Res. 1980;32:1–74. doi: 10.1016/s0065-230x(08)60360-7. [DOI] [PubMed] [Google Scholar]
  6. Diamond L., O'Brien T. G., Rovera G. Inhibition of adipose conversion of 3T3 fibroblasts by tumour promoters. Nature. 1977 Sep 15;269(5625):247–249. doi: 10.1038/269247a0. [DOI] [PubMed] [Google Scholar]
  7. Dicker P., Rozengurt E. Phorbol ester stimulation of Na influx and Na-K pump activity in Swiss 3T3 cells. Biochem Biophys Res Commun. 1981 May 15;100(1):433–441. doi: 10.1016/s0006-291x(81)80115-5. [DOI] [PubMed] [Google Scholar]
  8. Dicker P., Rozengurt E. Stimulation of DNA synthesis by tumour promoter and pure mitogenic factors. Nature. 1978 Dec 14;276(5689):723–726. doi: 10.1038/276723a0. [DOI] [PubMed] [Google Scholar]
  9. Driedger P. E., Blumberg P. M. Specific binding of phorbol ester tumor promoters. Proc Natl Acad Sci U S A. 1980 Jan;77(1):567–571. doi: 10.1073/pnas.77.1.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Driedger P. E., Blumberg P. M. The effect of phorbol diesters on chicken embryo fibroblasts. Cancer Res. 1977 Sep;37(9):3257–3265. [PubMed] [Google Scholar]
  11. Frantz C. N., Stiles C. D., Scher C. D. The tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate enhances the proliferative response of Balb/c-3T3 cells to hormonal growth factors. J Cell Physiol. 1979 Sep;100(3):413–424. doi: 10.1002/jcp.1041000305. [DOI] [PubMed] [Google Scholar]
  12. Gargus J. J., Slayman C. W. Mechanism and role of furosemide-sensitive K+ transport in L cells: a genetic approach. J Membr Biol. 1980;52(3):245–256. doi: 10.1007/BF01869193. [DOI] [PubMed] [Google Scholar]
  13. Geck P., Pietrzyk C., Burckhardt B. C., Pfeiffer B., Heinz E. Electrically silent cotransport on Na+, K+ and Cl- in Ehrlich cells. Biochim Biophys Acta. 1980 Aug 4;600(2):432–447. doi: 10.1016/0005-2736(80)90446-0. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. McRoberts J. A., Erlinger S., Rindler M. J., Saier M. H., Jr Furosemide-sensitive salt transport in the Madin-Darby canine kidney cell line. Evidence for the cotransport of Na+, K+, and Cl-. J Biol Chem. 1982 Mar 10;257(5):2260–2266. [PubMed] [Google Scholar]
  16. Mendoza S. A., Wigglesworth N. M., Pohjanpelto P., Rozengurt E. Na entry and Na-K pump activity in murine, hamster, and human cells--effect of monensin, serum, platelet extract, and viral transformation. J Cell Physiol. 1980 Apr;103(1):17–27. doi: 10.1002/jcp.1041030104. [DOI] [PubMed] [Google Scholar]
  17. Moroney J., Smith A., Tomei L. D., Wenner C. E. Stimulation of 86Rb+ and 32Pi movements in 3T3 cells by prostaglandins and phorbol esters. J Cell Physiol. 1978 Jun;95(3):287–294. doi: 10.1002/jcp.1040950306. [DOI] [PubMed] [Google Scholar]
  18. O'Brien T. G. Hexose transport in undifferentiated and differentiated BALB/c 3T3 preadipose cells: effects 12-O-tetradecanoylphorbol-13-acetate and insulin. J Cell Physiol. 1982 Jan;110(1):63–71. doi: 10.1002/jcp.1041100111. [DOI] [PubMed] [Google Scholar]
  19. O'Brien T. G., Lewis M. A., Diamond L. Ornithine decarboxylase activity and DNA synthesis after treatment of cells in culture with 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1979 Nov;39(11):4477–4480. [PubMed] [Google Scholar]
  20. O'Brien T. G., Saladik D., Diamond L. The tumor promoter 12-O-tetradecanoylphorbol-13-acetate stimulates lactate production in BALB/c 3T3 preadipose cells. Biochem Biophys Res Commun. 1979 May 14;88(1):103–110. doi: 10.1016/0006-291x(79)91702-9. [DOI] [PubMed] [Google Scholar]
  21. Plagemann P. G., Estensen R. D. Lack of correlation between effects of tumor promoter TPA on plasminogen activator production, phosphatidyl choline synthesis, and hexose transport in mammalian cell culture systems. J Cell Physiol. 1980 Jul;104(1):105–119. doi: 10.1002/jcp.1041040114. [DOI] [PubMed] [Google Scholar]
  22. Rindler M. J., McRoberts J. A., Saier M. H., Jr (Na+,K+)-cotransport in the Madin-Darby canine kidney cell line. Kinetic characterization of the interaction between Na+ and K+. J Biol Chem. 1982 Mar 10;257(5):2254–2259. [PubMed] [Google Scholar]
  23. Rozengurt E., Heppel L. A. Serum rapidly stimulates ouabain-sensitive 86-RB+ influx in quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4492–4495. doi: 10.1073/pnas.72.11.4492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schimmel S. D., Hallam T. Rapid alteration in Ca++ content and fluxes in phorbol 12-myristate 13-acetate treated myoblasts. Biochem Biophys Res Commun. 1980 Jan 29;92(2):624–630. doi: 10.1016/0006-291x(80)90379-4. [DOI] [PubMed] [Google Scholar]
  25. Schmidt W. F., 3rd, McManus T. J. Ouabain-insensitive salt and water movements in duck red cells. I. Kinetics of cation transport under hypertonic conditions. J Gen Physiol. 1977 Jul;70(1):59–79. doi: 10.1085/jgp.70.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schuldiner S., Rozengurt E. Na+/H+ antiport in Swiss 3T3 cells: mitogenic stimulation leads to cytoplasmic alkalinization. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7778–7782. doi: 10.1073/pnas.79.24.7778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shoyab M., Todaro G. J. Specific high affinity cell membrane receptors for biologically active phorbol and ingenol esters. Nature. 1980 Dec 4;288(5790):451–455. doi: 10.1038/288451a0. [DOI] [PubMed] [Google Scholar]
  28. Smith J. B., Rozengurt E. Serum stimulates the Na+,K+ pump in quiescent fibroblasts by increasing Na+ entry. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5560–5564. doi: 10.1073/pnas.75.11.5560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Spaggiare S., Wallach M. J., Tupper J. T. Potassium transport in normal and transformed mouse 3T3 cells. J Cell Physiol. 1976 Nov;89(3):403–416. doi: 10.1002/jcp.1040890306. [DOI] [PubMed] [Google Scholar]
  30. Wiley J. S., Cooper R. A. A furosemide-sensitive cotransport of sodium plus potassium in the human red cell. J Clin Invest. 1974 Mar;53(3):745–755. doi: 10.1172/JCI107613. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES