SpringerLink
Log in

Further studies on the charge-related alterations of methotrexate transport in Ehrlich ascites tumor cells by ionic liposomes: Correlation with liposome-cell association

  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Interaction of positively (phosphatidylcholine/stearylamine 5∶1) or negatively (phosphatidylcholine/stearic acid 5∶1) charged liposomes with Ehrlich ascites tumor cells for 1–5 min increases or decreases, respectively, the bidirectional fluxes of the folic acid analog, methotrexate. These effects on influx and efflux appear to be symmetrical since the liposomes do not change the intracellular level of methotrexate at the steady state. Influx kinetics show that these alterations result from an increase or decrease in theV max with no change in theK inm . These effects appear to be specific for the methotrexate-tetrahydrofolate carrier system since the transport of other compounds which utilize this carrier, aminopterin, 5-methyltetrahydrofolate, and 5-formyltetrahydrofolate, is affected similarly to methotrexate, whereas, the transport of folic acid, a compound similar in structure and charge but not significantly transported by this carrier is unaffected by liposomes. Once cells are exposed to charged liposomes, the effects on methotrexate transport cannot be reversed by washing the cells free of the extracellular liposomes. If, however, cells are exposed to liposomes of one charge, washed and then exposed to liposomes of the opposite charge, methotrexate influx is reversed to control rates. The effects of charged liposomes on methotrexate influx were not abolished by treating the cells with neuraminidase, metabolic inhibitors or lowering the temperature to 4°C. Studies on the uptake of [14C] liposomes show that these effects are not proportional to the total amount of lipid associated with the cell but result from an initial rapid liposome-cell association that is not dependent on temperature or energy metabolism nor related to cell surface charge.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Germany)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bartlett, G.R. 1959. Phosphorus assay in column chromatography.J. Biol. Chem. 234:466–468

    PubMed  Google Scholar 

  2. Bigon, E., Boarato, E., Bruni, A., Leon, A., Tofdano, D. 1979. Pharmacological effects of phosphatidylserine liposomes: Regulation of glycolysis and energy level in brain.Br. J. Pharmacol. 66:167–174

    PubMed  Google Scholar 

  3. Bobzien, W.F., III, Goldman, I.D. 1972. The mechanism of folate transport in rabbit reticulocytes.J. Clin. Invest. 51:1688–1696

    PubMed  Google Scholar 

  4. Bohn, B., Thies, C., Brossmer, R. 1977. Cell surface charge, sialic acid content and metabolic behavior of two tumor sublines. A comparative study.Eur. J. Cancer 13:1145–1150

    PubMed  Google Scholar 

  5. Bouma, S.R., Drislane, F.W., Huestis, W.H. 1977. Selective extraction of membrane bound proteins by phospholipid vesicles.J. Biol. Chem. 252:6759–6763

    PubMed  Google Scholar 

  6. Bruckdorfer, K.R., Edwards, P.A., Green, C. 1968. Properties of aqueous dispersions of phospholipid and cholesterol.Eur. J. Biochem. 4:506–511

    PubMed  Google Scholar 

  7. Burns, C.P., Luttenegger, D.G., Dudley, D.T., Buettner, G.R., Spector, A.A. 1979. Effect of modification of plasma membrane fatty acid composition on fluidity and methotrexate transport in L1210 murine leukemia cells.Cancer Res. 39:1726–1732

    PubMed  Google Scholar 

  8. Christie, W.W. 1973. Lipid Analysis. pp. 176–227. Pergamon Press, New York

    Google Scholar 

  9. Cooper, R.A. 1977. Abnormalities of cell-membrane fluidity in the pathogenesis of disease.N. Engl. J. Med. 297:371–377

    PubMed  Google Scholar 

  10. Dunnick, J.K., Rooke, J.D., Aragon, S., Kriss, J.P. 1976. Alteration of mammalian cells by interaction with artificial lipid vesicles.Cancer Res. 36:2385–2389

    PubMed  Google Scholar 

  11. Feinstein, M.B., Fernandez, S.M., Sha'afi, R.I. 1975. Fluidity of natural membranes and phosphatidylserine and ganglioside dispersions. Effects of anesthetics, cholesterol and proteinBiochim. Biophys. Acta 413:354–370

    PubMed  Google Scholar 

  12. Freise, J., Schmidt, F.W., Magerstedt, D. 1979. Effect of liposome-entrapped methotrexate on Ehrlich ascites tumor cells and uptake in primary liver cell tumor.J. Cancer Res. Clin. Oncol. 94:21–27

    PubMed  Google Scholar 

  13. Fry, D.W., White, J.C., Goldman, I.D. 1979. Alterations of the carrier-mediated transport of an anionic solute, methotrexate, by charged liposomes in Ehrlich ascites tumor cells.J. Membrane Biol. 50:123–140

    Google Scholar 

  14. Fry, D.W., White, J.C., Goldman, I.D. 1980. Effects of 2,4-dinitrophenol and other metabolic inhibitors on the bidirectional carrier fluxes, net transport, and intracellular binding of methotrexate in Ehrlich ascites tumor cells.Cancer Res. 40:3669–3673

    PubMed  Google Scholar 

  15. Goldman, I.D. 1969. Transport energetics of the folic acid analogue, methotrexate, in L1210 leukemia cells.J. Biol. Chem. 244:3779–3785

    PubMed  Google Scholar 

  16. Goldman, I.D. 1971. The characteristics of the membrane transport of amethopterin and the naturally occurring folates.Ann. N.Y. Acad. Sci. 186:400–422

    PubMed  Google Scholar 

  17. Goldman, I.D., Lichtenstein, N.S., Oliverio, V.T. 1968. Carriermediated transport of the folic acid analogue, methotrexate, in the L1210 leukemia cell.J. Biol. Chem. 243:5007–5017

    PubMed  Google Scholar 

  18. Huestis, W.H. 1977. A sodium-specific membrane permeability defect induced by phospholipid vesicles treatment of erythrocytes.J. Biol. Chem. 252:6764

    PubMed  Google Scholar 

  19. Huunan-Seppala, A. 1971. Cation permeability induced by spermine and polybrene in rat liver mitochondria.Bioenergetics 2:197–207

    Google Scholar 

  20. Inbar, M., Shinitszky, M. 1974. Increase of cholesterol level in the surface membrane of lymphoma cells and its inhibitory effect on ascites tumor development.Proc. Natl. Acad. Sci. USA 71:2128–2130

    PubMed  Google Scholar 

  21. Jackson, R.C., Niethammer D., Huennekens, F.M. 1975. Enzymic and transport mechanisms of amethopterin resistance in L1210 mouse leukemia cells.Cancer Biochem. Biophys. 1:151–155

    Google Scholar 

  22. Katchalski, E., Silman, I., Goldman, R. 1971. Effect of the microenvironment on the mode of action of immobilized enzymes.Adv. Enzymol. 34:445–536

    PubMed  Google Scholar 

  23. Kimelberg, H.K. 1976. Differential distribution of liposome-entrapped [3H] methotrexate and labeled lipids after intravenous injection in a primate.Biochim. Biophys. Acta 448:531–550

    PubMed  Google Scholar 

  24. Kimelberg, H.K., Mayhew, E.G. 1979. Properties and biological effects of liposomes and their uses in pharmacology and toxicity.CRC Crit. Rev. Toxicol. 6:25–79

    Google Scholar 

  25. Kimelberg, H.K., Tracy, T.F., Jr., Biddlecome, S.M., Bourke, R.S. 1976. The effect of entrapment in liposomes on the in vivo distribution of [3H] methotrexate in a primate.Cancer Res. 36:2949–2957

    PubMed  Google Scholar 

  26. Kinsky, S.C., Nicolotti, R.A. 1977. Immunological properties of model membranes.Annu. Rev. Biochem. 46:49–67

    PubMed  Google Scholar 

  27. Klein, I., Moore, L., Pastan, I. 1978. Effect of liposomes containing cholesterol on adenylate cyclase activity of cultured mammalian fibroblasts.Biochim. Biophys. Acta 506:42–53

    PubMed  Google Scholar 

  28. Kosloski, M.J., Rosen, F., Milholland, R., Papahadjopoulos, D. 1978. Effect of lipid vesicle (liposome) encapsulation of methotrexate on its chemotherapeutic efficacy in solid rodent tumors.Cancer Res. 38:2848–2853

    PubMed  Google Scholar 

  29. Meisner, H., Palmieri, F., Quagliariello, E. 1972. Effect of cations and protons on the kinetics of substrate uptake in rat liver mitochondria.Biochemistry 11:949–955

    PubMed  Google Scholar 

  30. Ozato, K., Huang, L., Pagano, R.E., 1978. Interactions of phospholipid vesicles with murine lymphocytes. II. Correlation between altered surface properties and enhanced proliferation response.Membr. Biochem. 1:27–42

    PubMed  Google Scholar 

  31. Pagano, R.E., Weinstein, J.N. 1978. Interaction of liposomes with mammalian cells.Annu. Rev. Biophys. Bioeng. 7:435–468

    PubMed  Google Scholar 

  32. Panzer, E.A., Jansons, V.K. 1979 Control of in vitro cytotoxity of positively charged liposomes.J. Cancer Res. Clin. Oncol. 95:29–37

    PubMed  Google Scholar 

  33. Papa, S., Guerrieri, F., Simone, S., Lorusso, M. 1972. Action of local anaesthetics on passive and energy-linked ion translocation in the inner mitochondrial membrane.Bioenergetics 3:553–568

    Google Scholar 

  34. Papahadjopoulos, D., Hui, S., Vail, W.J., Poste, G. 1976. Studies on membrane fusion: I. Interactions of pure phospholipid membranes and the effect of myristic acid, lysolecithin, proteins and dimethylsulfoxide.Biochim. Biophys. Acta 448:245–264

    Google Scholar 

  35. Papahadjopoulos, D., Jacobson, K., Poste, G., Shepherd, G. 1975. Effect of local anesthetics on membrane properties. I. Changes in the fluidity of phospholipid bilayers.Biochim. Biophys. Acta 394:504–519

    PubMed  Google Scholar 

  36. Poe, M. 1977. Acidic dissociation constants of folic acid, dihydrofolic acid and methotrexate.J. Biol. Chem. 252:3724–3728

    PubMed  Google Scholar 

  37. Poste, G., Allison, A.C. 1973. Membrane fusion.Biochim. Biophys. Acta 300:421–465

    PubMed  Google Scholar 

  38. Poste, G., Papahadjopoulos, D., Vail, W.J. 1976. Lipid vesicles as carriers for introducing biologically active materials into cells.Methods Cell Biol. 14:33–71

    PubMed  Google Scholar 

  39. Poznansky, M., Kirkwood, D., Solomon, A.K. 1973. Modulation of red cell K+ transport by membrane lipids.Biochim. Biophys. Acta 330:351–355

    PubMed  Google Scholar 

  40. Quagliariello, E., Palmieri, F., Drezioso, G. 1969. Kinetics of succinate uptake by rat-liver mitochondria.FEBS Lett. 4:251–254

    PubMed  Google Scholar 

  41. Rader, J.I., Niethammer, D., Huennekens, F.M. 1974. Effects of sulfhydryl inhibitors upon transport of folate compounds into L1210 cells.Biochem. Pharmacol. 23:2057–2059

    PubMed  Google Scholar 

  42. Rivnay, B., Globerson, A., Shinitzky, M. 1978 Perturbation of lymphocyte response to concanavalin A.Eur. J. Immunol. 8:185–189

    PubMed  Google Scholar 

  43. Shinitzky, M., Inbar, M. 1974. Difference in microviscosity induced by different cholesterol levels in the surface membrane layer of normal lymphocytes and malignant lymphoma cells.J. Mol. Biol. 85:603–615

    PubMed  Google Scholar 

  44. Singleton, W.S., Gray, M.S., Brown M.L., White, J.L. 1965. Chromatographically homogeneous lecithin from egg phospholipid.J. Am. Oil. Chem. Soc. 42:53–56

    PubMed  Google Scholar 

  45. Stoffel, W. 1975. Chemical synthesis of choline-labeled lecithins.In: Methods in Enzymology. S.P. Colowick and N.O. Kaplan, editors. Vol. 35, pp. 553–541. Academic Press, New York

    Google Scholar 

  46. Todd, J.A., Levine, A.M., Tokes, Z.A. 1980. Liposome-encapsulated methotrexate interactions with human chronic lymphocytic leukemia cells.J. Natl. Cancer Inst. 64:715–719

    PubMed  Google Scholar 

  47. Trauble, H., Eibl, H. 1974. Electrostatic effects on lipid phase transitions: Membrane structure and ionic environmentProc. Natl. Acad. Sci. USA 71:214–219

    PubMed  Google Scholar 

  48. Warren, L. 1959. The thiobarbituric acid assay of sialic acids.J. Biol. Chem. 234:1971–1975

    PubMed  Google Scholar 

  49. Weinstein, J.N., Magin, R.L., Cysyk, R.L., Zaharko, D.S. 1980. Treatment of solid L1210 murine tumors with local hyperthermia and temperature-sensitive liposomes containing methotrexate.Cancer Res. 40:1388–1395

    PubMed  Google Scholar 

  50. Weinstein, J.N., Magin, R.L., Yatvin, M.B., Zaharko, D.S. 1979. Liposomes and local hyperthermia: Selective delivery of methotrexate to heated tumors.Science 204:188–191

    PubMed  Google Scholar 

  51. Weiss, L., Zeigel, R.J. 1972. Heterogeneity of anionic sites at the electrokinetic surfaces of fixed Ehrlich ascites tumor cells.J. Theor. Biol. 34:21–27

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Medicine, Medical College of Virginia, 23298, Richmond, Virginia

    D. W. Fry & I. D. Goldman

Authors
  1. D. W. Fry
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I. D. Goldman
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fry, D.W., Goldman, I.D. Further studies on the charge-related alterations of methotrexate transport in Ehrlich ascites tumor cells by ionic liposomes: Correlation with liposome-cell association. J. Membrain Biol. 66, 87–95 (1982). https://doi.org/10.1007/BF01868485

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01868485

Key words

Search

Navigation