Skip to main content
Log in

Manganese uptake and release by cultured human hepatocarcinoma (Hep-G2) cells

  • Original Articles
  • Published:
Biological Trace Element Research Aims and scope Submit manuscript

Abstract

The liver is the primary organ involved in manganese (Mn) homeostasis. The human hepato-carcinoma cell line, Hep-G2, shows many liver specific functions. Consequently, Hep-G2 cells were investigated as a possible model of hepatic metabolism of Mn. Initial experiments showed that the concentration of Mn in the diet, or culture medium, similarly affected the retention of Mn by isolated rat hepatocytes and Hep-G2 cells. Manganese uptake by Hep-G2 cells suggested that uptake was followed by release from the cell. Uptake was saturable and half-maximal at 2.0 μmol Mn/L, and was inhibited by iodoacetate, vanadate, cold, and bepridil. The cations Fe2+, Cu2+, Ni2+, Cd2+, and Zn2+ decreased Mn uptake. Uptake was dependent on Calcium (Ca) concentration in a manner that resembled saturation kinetics. Cells that were pulsed with54Mn and then placed into nonradioactive medium quickly released a large portion of their internalized Mn. Release of internalized Mn could be inhibited by low temperature, nocodozole, quinacrine and sodium azide. These data show that Hep-G2 cells are a potentially good model of hepatic Mn metabolism. Mn is taken up by a facilitated process that may be related to Ca uptake. Release apparently is an active, controlled process, that may involve microtubules and lysosomes.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. L. Hurley and C. Keen, Manganese, in:Trace Elements in Human and Animal Nutrition, vol. 5, W. Mertz, ed., Academic, San Diego, CA, pp. 185–224 (1987).

    Google Scholar 

  2. D. Lee and P. Johnson,54Mn absorption and excretion in rats fed soy protein and casein diets (42852),Proc. Soc. Exp. Biol. Med. 190, 211–216 (1989).

    PubMed  CAS  Google Scholar 

  3. D. Greenberg, D. Copp, and E. Cuthbertson, Studies in mineral metabolism with the aid of artificial radioactive isotopes VII. The distribution and excretion, particularly by way of the bile, of iron, cobalt, and manganese,J. Biol. Chem. 147, 749–756 (1943).

    CAS  Google Scholar 

  4. C. Klassen, Biliary excretion of manganese in rats, rabbits and dogs,Toxicol. Appl. Pharmacol.29, 458–468 (1974).

    Article  Google Scholar 

  5. J. Finley, J. Caton, Z. Zhou, and K. Davison, A surgical model for determination of the true absorption and biliary excretion of manganese in concious swine.J. Nutr. 127, 2334–2341 (1997).

    PubMed  CAS  Google Scholar 

  6. B. Knowles, C. Howe, and D. Aden, Human hepatocellular carcinoma cell lines secrete the major plasma proteins and Hepatitis B surface antigen,Science 209, 497–499 (1980).

    Article  PubMed  CAS  Google Scholar 

  7. L. Klevay, H. Petering, and K. Stemmer, A controlled environment for trace metal experiments on animals,Environ. Sci. Technol. 5, 1196–1199 (1971).

    Article  CAS  Google Scholar 

  8. M. Berry and D. Friend, High yield preparation of isolated rat liver parenchymal cells: A biochemical and fine structural study,Cell Biol. 43, 506–520 (1969).

    Article  CAS  Google Scholar 

  9. R. Pilz, R. Willis, and J. Seegmiller, Regulation of human lymphoblast plasma membrane 5’-nucleotidase by zinc,J. Biol. Chem. 257, 13, 544–13,549 (1982).

    Google Scholar 

  10. J. Finley, P. Reeves, M. Briske-Anderson, and L. Johnson, Zinc uptake and transcellular movement by CACO-2 cells: Studies with media containing fetal bovine serum,J. Nutr Biochem. 6, 137–144 (1995).

    Article  Google Scholar 

  11. T. Chen, In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain,Exp. Cell Res. 104, 255–262 (1977).

    Article  PubMed  CAS  Google Scholar 

  12. M. Aschner and M. Gannon, Manganese (Mn) transport across the rat blood-brain barrier: saturable and transferrin dependent transport mechanisms,Brain Res. Bull.33, 345–349 (1994).

    Article  PubMed  CAS  Google Scholar 

  13. National Research Council, Recommended Dietary Allowances, National Academy Press, Washington, DC, (1989).

    Google Scholar 

  14. T. Galeotti, G. Palombini, and G. D. V. van Rossum, Manganese content and high affinity transport in liver and hepatoma,Arch. Biochem. Biophys. 322, 453–459 (1995).

    Article  PubMed  CAS  Google Scholar 

  15. M. Brandt and V. Schramm, Mammalian manganese metabolism and manganese uptake and distribution in rat hepatocytes, in:Manganese in metabolism and enzyme function, V. Schramm and F. Wedler, eds., Academic, Orlando, FL, pp. 3–16 (1986).

    Google Scholar 

  16. M. Frame and M. Milanick, Mn and Cd transport by the Na-Ca exchanger of ferret red blood cells,Am. J. Physiol. 261, 467–475 (1991).

    Google Scholar 

  17. S. Hagiwara and L. Byerly, Membrane biophysics of calcium currents,FASEB J. 40, 2220–2225 (1981).

    CAS  Google Scholar 

  18. J. Crofts and J. Barritt, The liver cell plasma membrane Ca2+ inflow systems exhibit a broad specificity for divalent metal cations,Biochem. J. 269, 579–587 (1990).

    PubMed  CAS  Google Scholar 

  19. R. D. Raffaniello, S. Lee, S. Teichberg, and R. A. Wapnir, Distinct mechanisms of zinc uptake at the apical and basolateral membranes of Caco-2 cells,J. Cell. Physiol. 152, 356–361 (1992).

    Article  PubMed  CAS  Google Scholar 

  20. R. Sormunen, S. Eskelinen, and V. Lehto, Bile caniliculus formation in cultured HepG2 cells,Lab. Invest. 68, 652–662 (1993).

    PubMed  CAS  Google Scholar 

  21. N. Javitt, R. Pfeffer, E. Kok, S. Burstein, B. Cohen, and K. Budai, Bile acid synthesis in cell culture,J. Biol. Chem. 264, 10384–10387 (1989).

    PubMed  CAS  Google Scholar 

  22. W. Mertz, Trace elements in human and animal nutrition, in:Trace Elements in Human and Animal Nutrition, W. Mertz, ed., Academic, NY (1985).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

The U.S. Department of Agriculture, Agricultural Research Service, Northern Plains Area, is an equal opportunity/affirmative action employer and all agency services are available without discrimination.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fimley, J.W. Manganese uptake and release by cultured human hepatocarcinoma (Hep-G2) cells. Biol Trace Elem Res 64, 101–118 (1998). https://doi.org/10.1007/BF02783328

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02783328

Index entries

Navigation