Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1992 Jan;140(1):45–55.

Effects of transforming growth factor-beta on collagen synthesis by normal rat kidney epithelial cells.

J J Creely 1, S J DiMari 1, A M Howe 1, M A Haralson 1
PMCID: PMC1886252  PMID: 1731530

Abstract

The effects of transforming growth factor-beta (TGF-beta) on the growth of and collagen production by NRK52E cells, a clonal line established from normal rat kidney epithelial cells, have been characterized. NRK52E cells were grown in the absence or presence of TGF-beta for 4 days followed by incubation for 24 hours in serum-free medium containing [3H]proline. The secreted and cell-associated collagens produced by control and experimental cultures were isolated by limited pepsin digestion and differential salt fractionation. TGF-beta inhibited proliferation by about 50% but did not affect overall culture morphology. Both protein and collagen synthesis were increased in experimental cultures, but the increase in total collagen production exceeded that of total protein synthesis. Although NRK52E cells grown in the presence of TGF-beta continued to produce the same types of collagen (types I, III, IV, and V), their relative amounts were changed. In the experimental cultures, type I collagen production was increased eightfold, types III and V collagen levels were increased two-fold, but type IV production was only slightly enhanced. In addition to increasing total collagen production by about fivefold, TGF-beta increased the ratio of type I to type III about threefold but minimally affected the ratio of secreted to cell-associated molecules. These findings establish that TGF-beta specifically affects collagen production in NRK52E cells and that these alterations differ in many ways from the affects of epidermal growth factor. Because TGF-beta increased total collagen expression, these results provide additional evidence implicating this growth factor as a positive mediator of matrix accumulation in renal disease.

Full text

PDF
46

Images in this article

47Figure 1
on p.47

Selected References

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

  1. Ardaillou N., Bellon G., Nivez M. P., Rakotoarison S., Ardaillou R. Quantification of collagen synthesis by cultured human glomerular cells. Biochim Biophys Acta. 1989 Jun 27;991(3):445–452. doi: 10.1016/0304-4165(89)90071-8. [DOI] [PubMed] [Google Scholar]
  2. Assoian R. K., Grotendorst G. R., Miller D. M., Sporn M. B. Cellular transformation by coordinated action of three peptide growth factors from human platelets. 1984 Jun 28-Jul 4Nature. 309(5971):804–806. doi: 10.1038/309804a0. [DOI] [PubMed] [Google Scholar]
  3. Assoian R. K., Komoriya A., Meyers C. A., Miller D. M., Sporn M. B. Transforming growth factor-beta in human platelets. Identification of a major storage site, purification, and characterization. J Biol Chem. 1983 Jun 10;258(11):7155–7160. [PubMed] [Google Scholar]
  4. Barnard J. A., Lyons R. M., Moses H. L. The cell biology of transforming growth factor beta. Biochim Biophys Acta. 1990 Jun 1;1032(1):79–87. doi: 10.1016/0304-419x(90)90013-q. [DOI] [PubMed] [Google Scholar]
  5. Border W. A., Okuda S., Languino L. R., Ruoslahti E. Transforming growth factor-beta regulates production of proteoglycans by mesangial cells. Kidney Int. 1990 Feb;37(2):689–695. doi: 10.1038/ki.1990.35. [DOI] [PubMed] [Google Scholar]
  6. Border W. A., Okuda S., Languino L. R., Sporn M. B., Ruoslahti E. Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1. Nature. 1990 Jul 26;346(6282):371–374. doi: 10.1038/346371a0. [DOI] [PubMed] [Google Scholar]
  7. Bruijn J. A., Hogendoorn P. C., Hoedemaeker P. J., Fleuren G. J. The extracellular matrix in pathology. J Lab Clin Med. 1988 Feb;111(2):140–149. [PubMed] [Google Scholar]
  8. Coimbra T., Wiggins R., Noh J. W., Merritt S., Phan S. H. Transforming growth factor-beta production in anti-glomerular basement membrane disease in the rabbit. Am J Pathol. 1991 Jan;138(1):223–234. [PMC free article] [PubMed] [Google Scholar]
  9. Coustry F., Gillery P., Maquart F. X., Borel J. P. Effect of transforming growth factor beta on fibroblasts in three-dimensional lattice cultures. FEBS Lett. 1990 Mar 26;262(2):339–341. doi: 10.1016/0014-5793(90)80223-6. [DOI] [PubMed] [Google Scholar]
  10. Creely J. J., Commers P. A., Haralson M. A. Synthesis of type III collagen by cultured kidney epithelial cells. Connect Tissue Res. 1988;18(2):107–122. doi: 10.3109/03008208809008063. [DOI] [PubMed] [Google Scholar]
  11. Creely J. J., DiMari S. J., Howe A. M., Hyde C. P., Haralson M. A. Effects of epidermal growth factor on collagen synthesis by an epithelioid cell line derived from normal rat kidney. Am J Pathol. 1990 Jun;136(6):1247–1257. [PMC free article] [PubMed] [Google Scholar]
  12. DiMari S. J., Howe A. M., Haralson M. A. Effects of transforming growth factor-beta on collagen synthesis by fetal rat lung epithelial cells. Am J Respir Cell Mol Biol. 1991 May;4(5):455–462. doi: 10.1165/ajrcmb/4.5.455. [DOI] [PubMed] [Google Scholar]
  13. Dubois C. H., Foidart J. B., Hautier M. B., Dechenne C. A., Lemaire M. J., Mahieu P. R. Proliferative glomerulonephritis in rats: evidence that mononuclear phagocytes infiltrating the glomeruli stimulate the proliferation of endothelial and mesangial cells. Eur J Clin Invest. 1981 Apr;11(2 Suppl 1):91–104. doi: 10.1111/j.1365-2362.1981.tb02045.x. [DOI] [PubMed] [Google Scholar]
  14. Federspiel S. J., DiMari S. J., Guerry-Force M. L., Haralson M. A. Extracellular matrix biosynthesis by cultured fetal rat lung epithelial cells. II. Effects of acute exposure to epidermal growth factor and retinoic acid on collagen biosynthesis. Lab Invest. 1990 Oct;63(4):455–466. [PubMed] [Google Scholar]
  15. Fine A., Goldstein R. H. The effect of transforming growth factor-beta on cell proliferation and collagen formation by lung fibroblasts. J Biol Chem. 1987 Mar 15;262(8):3897–3902. [PubMed] [Google Scholar]
  16. Graham M. F., Bryson G. R., Diegelmann R. F. Transforming growth factor beta 1 selectively augments collagen synthesis by human intestinal smooth muscle cells. Gastroenterology. 1990 Aug;99(2):447–453. doi: 10.1016/0016-5085(90)91028-5. [DOI] [PubMed] [Google Scholar]
  17. Haralson M. A., Federspiel S. J., Martinez-Hernandez A., Rhodes R. K., Miller E. J. Synthesis of [pro alpha 1(IV)]3 collagen molecules by cultured embryo-derived parietal yolk sac cells. Biochemistry. 1985 Oct 8;24(21):5792–5797. doi: 10.1021/bi00342a016. [DOI] [PubMed] [Google Scholar]
  18. Haralson M. A., Jacobson H. R., Hoover R. L. Collagen polymorphism in cultured rat kidney mesangial cells. Lab Invest. 1987 Nov;57(5):513–523. [PubMed] [Google Scholar]
  19. Haverty T. P., Kelly C. J., Hines W. H., Amenta P. S., Watanabe M., Harper R. A., Kefalides N. A., Neilson E. G. Characterization of a renal tubular epithelial cell line which secretes the autologous target antigen of autoimmune experimental interstitial nephritis. J Cell Biol. 1988 Oct;107(4):1359–1368. doi: 10.1083/jcb.107.4.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Holley R. W., Böhlen P., Fava R., Baldwin J. H., Kleeman G., Armour R. Purification of kidney epithelial cell growth inhibitors. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5989–5992. doi: 10.1073/pnas.77.10.5989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Howe P. H., Cunningham M. R., Leof E. B. Distinct pathways regulate transforming growth factor beta 1-stimulated proto-oncogene and extracellular matrix gene expression. J Cell Physiol. 1990 Jan;142(1):39–45. doi: 10.1002/jcp.1041420106. [DOI] [PubMed] [Google Scholar]
  22. Humes H. D., Beals T. F., Cieslinski D. A., Sanchez I. O., Page T. P. Effects of transforming growth factor-beta, transforming growth factor-alpha, and other growth factors on renal proximal tubule cells. Lab Invest. 1991 Apr;64(4):538–545. [PubMed] [Google Scholar]
  23. Ignotz R. A., Massagué J. Transforming growth factor-beta stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J Biol Chem. 1986 Mar 25;261(9):4337–4345. [PubMed] [Google Scholar]
  24. Kanda S., Nomata K., Saha P. K., Nishimura N., Yamada J., Kanatake H., Saito Y. Growth factor regulation of the renal cortical tubular cells by epidermal growth factor, insulin-like growth factor-I, acidic and basic fibroblast growth factor, and transforming growth factor-beta in serum free culture. Cell Biol Int Rep. 1989 Aug;13(8):687–699. doi: 10.1016/0309-1651(89)90045-3. [DOI] [PubMed] [Google Scholar]
  25. Killen P. D., Striker G. E. Human glomerular visceral epithelial cells synthesize a basal lamina collagen in vitro. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3518–3522. doi: 10.1073/pnas.76.7.3518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Klagsbrun M., Edelman E. R. Biological and biochemical properties of fibroblast growth factors. Implications for the pathogenesis of atherosclerosis. Arteriosclerosis. 1989 May-Jun;9(3):269–278. doi: 10.1161/01.atv.9.3.269. [DOI] [PubMed] [Google Scholar]
  27. Klahr S., Schreiner G., Ichikawa I. The progression of renal disease. N Engl J Med. 1988 Jun 23;318(25):1657–1666. doi: 10.1056/NEJM198806233182505. [DOI] [PubMed] [Google Scholar]
  28. Leheup B. P., Federspiel S. J., Guerry-Force M. L., Wetherall N. T., Commers P. A., DiMari S. J., Haralson M. A. Extracellular matrix biosynthesis by cultured fetal rat lung epithelial cells. I. Characterization of the clone and the major genetic types of collagen produced. Lab Invest. 1989 Jun;60(6):791–807. [PubMed] [Google Scholar]
  29. MacKay K., Striker L. J., Stauffer J. W., Doi T., Agodoa L. Y., Striker G. E. Transforming growth factor-beta. Murine glomerular receptors and responses of isolated glomerular cells. J Clin Invest. 1989 Apr;83(4):1160–1167. doi: 10.1172/JCI113996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Noda M., Rodan G. A. Type beta transforming growth factor regulates expression of genes encoding bone matrix proteins. Connect Tissue Res. 1989;21(1-4):71–75. doi: 10.3109/03008208909049997. [DOI] [PubMed] [Google Scholar]
  31. Nugent M. A., Newman M. J. Inhibition of normal rat kidney cell growth by transforming growth factor-beta is mediated by collagen. J Biol Chem. 1989 Oct 25;264(30):18060–18067. [PubMed] [Google Scholar]
  32. O'Keefe R. J., Puzas J. E., Brand J. S., Rosier R. N. Effects of transforming growth factor-beta on matrix synthesis by chick growth plate chondrocytes. Endocrinology. 1988 Jun;122(6):2953–2961. doi: 10.1210/endo-122-6-2953. [DOI] [PubMed] [Google Scholar]
  33. Okuda S., Languino L. R., Ruoslahti E., Border W. A. Elevated expression of transforming growth factor-beta and proteoglycan production in experimental glomerulonephritis. Possible role in expansion of the mesangial extracellular matrix. J Clin Invest. 1990 Aug;86(2):453–462. doi: 10.1172/JCI114731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Oomura A., Nakamura T., Arakawa M., Ooshima A., Isemura M. Alterations in the extracellular matrix components in human glomerular diseases. Virchows Arch A Pathol Anat Histopathol. 1989;415(2):151–159. doi: 10.1007/BF00784353. [DOI] [PubMed] [Google Scholar]
  35. Redini F., Galera P., Mauviel A., Loyau G., Pujol J. P. Transforming growth factor beta stimulates collagen and glycosaminoglycan biosynthesis in cultured rabbit articular chondrocytes. FEBS Lett. 1988 Jul 4;234(1):172–176. doi: 10.1016/0014-5793(88)81327-9. [DOI] [PubMed] [Google Scholar]
  36. Roberts A. B., Anzano M. A., Lamb L. C., Smith J. M., Sporn M. B. New class of transforming growth factors potentiated by epidermal growth factor: isolation from non-neoplastic tissues. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5339–5343. doi: 10.1073/pnas.78.9.5339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Roberts A. B., Flanders K. C., Kondaiah P., Thompson N. L., Van Obberghen-Schilling E., Wakefield L., Rossi P., de Crombrugghe B., Heine U., Sporn M. B. Transforming growth factor beta: biochemistry and roles in embryogenesis, tissue repair and remodeling, and carcinogenesis. Recent Prog Horm Res. 1988;44:157–197. doi: 10.1016/b978-0-12-571144-9.50010-7. [DOI] [PubMed] [Google Scholar]
  38. Roberts A. B., Heine U. I., Flanders K. C., Sporn M. B. Transforming growth factor-beta. Major role in regulation of extracellular matrix. Ann N Y Acad Sci. 1990;580:225–232. doi: 10.1111/j.1749-6632.1990.tb17931.x. [DOI] [PubMed] [Google Scholar]
  39. Roberts A. B., Sporn M. B., Assoian R. K., Smith J. M., Roche N. S., Wakefield L. M., Heine U. I., Liotta L. A., Falanga V., Kehrl J. H. Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4167–4171. doi: 10.1073/pnas.83.12.4167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rupard J. H., Dimari S. J., Damjanov I., Haralson M. A. Synthesis of type I homotrimer collagen molecules by cultured human lung adenocarcinoma cells. Am J Pathol. 1988 Nov;133(2):316–326. [PMC free article] [PubMed] [Google Scholar]
  41. Scheinman J. I., Brown D. M., Michael A. F. Collagen synthesis by human glomerular cells in culture. Biochim Biophys Acta. 1978 Aug 3;542(1):128–136. doi: 10.1016/0304-4165(78)90239-8. [DOI] [PubMed] [Google Scholar]
  42. Sporn M. B., Roberts A. B. Peptide growth factors and inflammation, tissue repair, and cancer. J Clin Invest. 1986 Aug;78(2):329–332. doi: 10.1172/JCI112580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Striker G. E., Striker L. J. Glomerular cell culture. Lab Invest. 1985 Aug;53(2):122–131. [PubMed] [Google Scholar]
  44. Striker L. J., Peten E. P., Elliot S. J., Doi T., Striker G. E. Mesangial cell turnover: effect of heparin and peptide growth factors. Lab Invest. 1991 Apr;64(4):446–456. [PubMed] [Google Scholar]
  45. Tan E. M., Dodge G. R., Sorger T., Kovalszky I., Unger G. A., Yang L., Levine E. M., Iozzo R. V. Modulation of extracellular matrix gene expression by heparin and endothelial cell growth factor in human smooth muscle cells. Lab Invest. 1991 Apr;64(4):474–482. [PubMed] [Google Scholar]
  46. Torbohm I., Berger B., Schönermark M., von Kempis J., Rother K., Hänsch G. M. Modulation of collagen synthesis in human glomerular epithelial cells by interleukin 1. Clin Exp Immunol. 1989 Mar;75(3):427–431. [PMC free article] [PubMed] [Google Scholar]
  47. Tucker R. F., Shipley G. D., Moses H. L., Holley R. W. Growth inhibitor from BSC-1 cells closely related to platelet type beta transforming growth factor. Science. 1984 Nov 9;226(4675):705–707. doi: 10.1126/science.6093254. [DOI] [PubMed] [Google Scholar]
  48. Wahl S. M., McCartney-Francis N., Allen J. B., Dougherty E. B., Dougherty S. F. Macrophage production of TGF-beta and regulation by TGF-beta. Ann N Y Acad Sci. 1990;593:188–196. doi: 10.1111/j.1749-6632.1990.tb16111.x. [DOI] [PubMed] [Google Scholar]
  49. Wrana J. L., Maeno M., Hawrylyshyn B., Yao K. L., Domenicucci C., Sodek J. Differential effects of transforming growth factor-beta on the synthesis of extracellular matrix proteins by normal fetal rat calvarial bone cell populations. J Cell Biol. 1988 Mar;106(3):915–924. doi: 10.1083/jcb.106.3.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wrana J. L., Sodek J., Ber R. L., Bellows C. G. The effects of platelet-derived transforming growth factor beta on normal human diploid gingival fibroblasts. Eur J Biochem. 1986 Aug 15;159(1):69–76. doi: 10.1111/j.1432-1033.1986.tb09834.x. [DOI] [PubMed] [Google Scholar]
  51. Yee J., Kuncio G. S., Neilson E. G. Tubulointerstitial injury following glomerulonephritis. Semin Nephrol. 1991 May;11(3):361–366. [PubMed] [Google Scholar]
  52. de Larco J. E., Todaro G. J. Epithelioid and fibroblastic rat kidney cell clones: epidermal growth factor (EGF) receptors and the effect of mouse sarcoma virus transformation. J Cell Physiol. 1978 Mar;94(3):335–342. doi: 10.1002/jcp.1040940311. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES