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. 1984 Feb 1;98(2):596–601. doi: 10.1083/jcb.98.2.596

A lipophilic iron chelator can replace transferrin as a stimulator of cell proliferation and differentiation

PMCID: PMC2113117  PMID: 6693498

Abstract

Of the different growth supplements used in chemically defined media, only transferrin is required for differentiation of tubules in the embryonic mouse metanephros. Since transferrin is an iron-carrying protein, we asked whether iron is crucial for tubulogenesis. Differentiation of metanephric tubules both in whole embryonic kidneys and in a transfilter system was studied. The tissues were grown in chemically defined media containing transferrin, apotransferrin, the metal-chelator complex ferric pyridoxal isonicotinoyl hydrazone (FePIH), and excesses of ferric ion. Although we found that apotransferrin was not as effective as iron-loaded transferrin in promoting proliferation in the differentiating kidneys, excess ferric ion at up to 100 microM, five times the normal serum concentration, could not promote differentiation or proliferation. However, iron coupled to the nonphysiological, lipophilic iron chelator, pyridoxal isonicotinoyl hydrazone, to form FePIH, could sustain levels of cell proliferation and tubulogenesis similar to those attained by transferrin. Thus, the role of transferrin in cell proliferation during tubulogenesis is solely to provide iron. Since FePIH apparently bypasses the receptor-mediated route of iron intake, the use of FePIH as a tool for investigating cell proliferation and its regulation is suggested.

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Selected References

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  1. Aisen P., Leibman A., Reich H. A. Studies on the binding of iron to transferrin and conalbumin. J Biol Chem. 1966 Apr 25;241(8):1666–1671. [PubMed] [Google Scholar]
  2. Barnes D., Sato G. Serum-free cell culture: a unifying approach. Cell. 1980 Dec;22(3):649–655. doi: 10.1016/0092-8674(80)90540-1. [DOI] [PubMed] [Google Scholar]
  3. Bates G. W., Schlabach M. R. The reaction of ferric salts with transferrin. J Biol Chem. 1973 May 10;248(9):3228–3232. [PubMed] [Google Scholar]
  4. Bleil J. D., Bretscher M. S. Transferrin receptor and its recycling in HeLa cells. EMBO J. 1982;1(3):351–355. doi: 10.1002/j.1460-2075.1982.tb01173.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Block T., Bothwell M. Use of iron- or selenium-coupled monoclonal antibodies to cell-surface antigens as a positive selection system for cells. Nature. 1983 Jan 27;301(5898):342–344. doi: 10.1038/301342a0. [DOI] [PubMed] [Google Scholar]
  6. Cikrt M., Ponka P., Necas E., Neuwirt J. Biliary iron excretion in rats following pyridoxal isonicotinoyl hydrazone. Br J Haematol. 1980 Jun;45(2):275–283. doi: 10.1111/j.1365-2141.1980.tb07147.x. [DOI] [PubMed] [Google Scholar]
  7. Dautry-Varsat A., Ciechanover A., Lodish H. F. pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2258–2262. doi: 10.1073/pnas.80.8.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ekblom P., Lehtonen E., Saxén L., Timpl R. Shift in collagen type as an early response to induction of the metanephric mesenchyme. J Cell Biol. 1981 May;89(2):276–283. doi: 10.1083/jcb.89.2.276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ekblom P., Thesleff I., Lehto V. P., Virtanen I. Distribution of the transferrin receptor in normal human fibroblasts and fibrosarcoma cells. Int J Cancer. 1983 Jan 15;31(1):111–117. doi: 10.1002/ijc.2910310118. [DOI] [PubMed] [Google Scholar]
  10. Ekblom P., Thesleff I., Miettinen A., Saxén L. Organogenesis in a defined medium supplemented with transferrin. Cell Differ. 1981 Nov;10(5):281–288. doi: 10.1016/0045-6039(81)90010-5. [DOI] [PubMed] [Google Scholar]
  11. Ekblom P., Thesleff I., Saxén L., Miettinen A., Timpl R. Transferrin as a fetal growth factor: acquisition of responsiveness related to embryonic induction. Proc Natl Acad Sci U S A. 1983 May;80(9):2651–2655. doi: 10.1073/pnas.80.9.2651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Enns C. A., Larrick J. W., Suomalainen H., Schroder J., Sussman H. H. Co-migration and internalization of transferrin and its receptor on K562 cells. J Cell Biol. 1983 Aug;97(2):579–585. doi: 10.1083/jcb.97.2.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fernandez-Pol J. A., Bono V. H., Jr, Johnson G. S. Control of growth by picolinic acid: differential response of normal and transformed cells. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2889–2893. doi: 10.1073/pnas.74.7.2889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. GROBSTEIN C. Trans-filter induction of tubules in mouse metanephrogenic mesenchyme. Exp Cell Res. 1956 Apr;10(2):424–440. doi: 10.1016/0014-4827(56)90016-7. [DOI] [PubMed] [Google Scholar]
  15. Hamilton T. A., Wada H. G., Sussman H. H. Identification of transferrin receptors on the surface of human cultured cells. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6406–6410. doi: 10.1073/pnas.76.12.6406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harding C., Heuser J., Stahl P. Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes. J Cell Biol. 1983 Aug;97(2):329–339. doi: 10.1083/jcb.97.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hopkins C. R., Trowbridge I. S. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J Cell Biol. 1983 Aug;97(2):508–521. doi: 10.1083/jcb.97.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Huebers H. A., Csiba E., Huebers E., Finch C. A. Competitive advantage of diferric transferrin in delivering iron to reticulocytes. Proc Natl Acad Sci U S A. 1983 Jan;80(1):300–304. doi: 10.1073/pnas.80.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Iacopetta B. J., Morgan E. H., Yeoh G. C. Transferrin receptors and iron uptake during erythroid cell development. Biochim Biophys Acta. 1982 May 7;687(2):204–210. doi: 10.1016/0005-2736(82)90547-8. [DOI] [PubMed] [Google Scholar]
  20. Ii I., Kimura I., Ozawa E. A myotrophic protein from chick embryo extract: its purification, identity to transferrin, and indispensability for avian myogenesis. Dev Biol. 1982 Dec;94(2):366–377. doi: 10.1016/0012-1606(82)90354-2. [DOI] [PubMed] [Google Scholar]
  21. JANDL J. H., INMAN J. K., SIMMONS R. L., ALLEN D. W. Transfer of iron from serum iron-binding protein to human reticulocytes. J Clin Invest. 1959 Jan 1;38(1 Pt 1):161–185. doi: 10.1172/JCI103786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Klausner R. D., Ashwell G., van Renswoude J., Harford J. B., Bridges K. R. Binding of apotransferrin to K562 cells: explanation of the transferrin cycle. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2263–2266. doi: 10.1073/pnas.80.8.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Martinez-Medellin J., Schulman H. M. The kinetics of iron and transferrin incorporation into rabbit erythroid cells and the nature of stromal-bound iron. Biochim Biophys Acta. 1972 Apr 21;264(2):272–274. doi: 10.1016/0304-4165(72)90291-7. [DOI] [PubMed] [Google Scholar]
  24. Nordling S., Aho S. Fluorimetric microassay of DNA using a modified thiobarbituric acid assay. Anal Biochem. 1981 Aug;115(2):260–266. doi: 10.1016/0003-2697(81)90004-x. [DOI] [PubMed] [Google Scholar]
  25. Nunez M. T., Cole E. S., Glass J. The reticulocyte plasma membrane pathway of iron uptake as determined by the mechanism of alpha, alpha'-dipyridyl inhibition. J Biol Chem. 1983 Jan 25;258(2):1146–1151. [PubMed] [Google Scholar]
  26. Octave J. N., Schneider Y. J., Hoffmann P., Trouet A., Crichton R. R. Transferrin uptake by cultured rat embryo fibroblasts. The influence of lysosomotropic agents, iron chelators and colchicine on the uptake of iron and transferrin. Eur J Biochem. 1982 Apr 1;123(2):235–240. doi: 10.1111/j.1432-1033.1982.tb19758.x. [DOI] [PubMed] [Google Scholar]
  27. Perez-Infante V., Mather J. P. The role of transferrin in the growth of testicular cell lines in serum-free medium. Exp Cell Res. 1982 Dec;142(2):325–332. doi: 10.1016/0014-4827(82)90374-3. [DOI] [PubMed] [Google Scholar]
  28. Ponka P., Borová J., Neuwirt J., Fuchs O. Mobilization of iron from reticulocytes. Identification of pyridoxal isonicotinoyl hydrazone as a new iron chelating agent. FEBS Lett. 1979 Jan 15;97(2):317–321. doi: 10.1016/0014-5793(79)80111-8. [DOI] [PubMed] [Google Scholar]
  29. Ponka P., Borová J., Neuwirt J., Fuchs O., Necas E. A study of intracellular iron metabolism using pyridoxal isonicotinoyl hydrazone and other synthetic chelating agents. Biochim Biophys Acta. 1979 Aug 22;586(2):278–297. doi: 10.1016/0304-4165(79)90100-4. [DOI] [PubMed] [Google Scholar]
  30. Ponka P., Schulman H. M., Wilczynska A. Ferric pyridoxal isonicotinoyl hydrazone can provide iron for heme synthesis in reticulocytes. Biochim Biophys Acta. 1982 Oct 8;718(2):151–156. doi: 10.1016/0304-4165(82)90213-6. [DOI] [PubMed] [Google Scholar]
  31. Richter A., Sanford K. K., Evans V. J. Influence of oxygen and culture media on plating efficiency of some mammalian tissue cells. J Natl Cancer Inst. 1972 Dec;49(6):1705–1712. doi: 10.1093/jnci/49.6.1705. [DOI] [PubMed] [Google Scholar]
  32. Rudland P. S., Durbin H., Clingan D., de Asua L. J. Iron salts and transferrin are specifically required for cell division of cultured 3T6 cells. Biochem Biophys Res Commun. 1977 Apr 11;75(3):556–562. doi: 10.1016/0006-291x(77)91508-x. [DOI] [PubMed] [Google Scholar]
  33. Saxén L., Koskimies O., Lahti A., Miettinen H., Rapola J., Wartiovaara J. Differentiation of kidney mesenchyme in an experimental model system. Adv Morphog. 1968;7:251–293. doi: 10.1016/b978-1-4831-9954-2.50011-2. [DOI] [PubMed] [Google Scholar]
  34. Saxén L., Lehtonen E. Transfilter induction of kidney tubules as a function of the extent and duration of intercellular contacts. J Embryol Exp Morphol. 1978 Oct;47:97–109. [PubMed] [Google Scholar]
  35. Shindelman J. E., Ortmeyer A. E., Sussman H. H. Demonstration of the transferrin receptor in human breast cancer tissue. Potential marker for identifying dividing cells. Int J Cancer. 1981 Mar 15;27(3):329–334. doi: 10.1002/ijc.2910270311. [DOI] [PubMed] [Google Scholar]
  36. Sibille J. C., Octave J. N., Schneider Y. J., Trouet A., Crichton R. R. Transferrin protein and iron uptake by cultured hepatocytes. FEBS Lett. 1982 Dec 27;150(2):365–369. doi: 10.1016/0014-5793(82)80769-2. [DOI] [PubMed] [Google Scholar]
  37. Sutherland R., Delia D., Schneider C., Newman R., Kemshead J., Greaves M. Ubiquitous cell-surface glycoprotein on tumor cells is proliferation-associated receptor for transferrin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4515–4519. doi: 10.1073/pnas.78.7.4515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Takahashi K., Tavassoli M. Biphasic uptake of iron-transferrin complex by L1210 murine leukemia cells and rat reticulocytes. Biochim Biophys Acta. 1982 Feb 8;685(1):6–12. doi: 10.1016/0005-2736(82)90027-x. [DOI] [PubMed] [Google Scholar]
  39. Tormey D. C., Imrie R. C., Mueller G. C. Identification of transferrin as a lymphocyte growth promoter in human serum. Exp Cell Res. 1972 Sep;74(1):163–169. doi: 10.1016/0014-4827(72)90492-2. [DOI] [PubMed] [Google Scholar]
  40. Trowbridge I. S., Lesley J., Schulte R. Murine cell surface transferrin receptor: studies with an anti-receptor monoclonal antibody. J Cell Physiol. 1982 Sep;112(3):403–410. doi: 10.1002/jcp.1041120314. [DOI] [PubMed] [Google Scholar]
  41. Trowbridge I. S., Lopez F. Monoclonal antibody to transferrin receptor blocks transferrin binding and inhibits human tumor cell growth in vitro. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1175–1179. doi: 10.1073/pnas.79.4.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Trowbridge I. S., Omary M. B. Human cell surface glycoprotein related to cell proliferation is the receptor for transferrin. Proc Natl Acad Sci U S A. 1981 May;78(5):3039–3043. doi: 10.1073/pnas.78.5.3039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Vogt A., Mishell R. I., Dutton R. W. Stimulation of DNA synthesis in cultures of mouse spleen cell suspensions by bovine transferrin. Exp Cell Res. 1969 Feb;54(2):195–200. doi: 10.1016/0014-4827(69)90232-8. [DOI] [PubMed] [Google Scholar]
  44. White G. P., Jacobs A. Iron uptake by Chang cells from transferrin, nitriloacetate and citrate complexes: the effects of iron-loading and chelation with desferrioxamine. Biochim Biophys Acta. 1978 Oct 3;543(2):217–225. doi: 10.1016/0304-4165(78)90066-1. [DOI] [PubMed] [Google Scholar]
  45. van Renswoude J., Bridges K. R., Harford J. B., Klausner R. D. Receptor-mediated endocytosis of transferrin and the uptake of fe in K562 cells: identification of a nonlysosomal acidic compartment. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6186–6190. doi: 10.1073/pnas.79.20.6186. [DOI] [PMC free article] [PubMed] [Google Scholar]

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