, Volume 34, Issue 5, pp 307–313

Binding and biological effects of insulin, insulin analogues and insulin-like growth factors in rat aortic smooth muscle cells. Comparison of maximal growth promoting activities

  • K. E. Bornfeldt
  • R. A. Gidlöf
  • A. Wasteson
  • M. Lake
  • A. Skottner
  • H. J. Arnqvist


Binding and growth promoting effects of insulin, insulin analogues modified in the B chain, proinsulin, insulin-like growth factor-I and -II were studied in cultured rat aortic smooth muscle cells. Specific binding of125I-insulin was 0.9±0.2% of total 125I-insulin added, and the IC50-value was estimated to 8.9 pmol/1. The insulin analogue B10 Asp tended to be more potent than insulin in displacing 125I-insulin, B28 Asp was equipotent, B9 Asp/B27 Glu was approximately 100 times less potent and insulin-like growth factor-I more than 1000 times less potent than insulin. Specific binding of 125I-insulin-like growth factor-I after 4 h incubation at 10 °C was five times higher than the specific binding of insulin (4.4±0.4% of total 125I-insulin-like growth factor-I added), and the IC50-value was 0.3 nmol/l. Insulin was approximately 500 times less potent than insulin-like growth factor-I in displacing 125I-insulin-like growth factor-I. The insulin analogue B10 Asp was slightly more potent and analogue B28 Asp was equipotent with insulin. Analogue B9 Asp/B27 Glu was ten times less potent and proinsulin was more than ten times less potent than insulin. The order of potency was similar for 3H-thymidine incorporation into DNA: insulin-like growth factor-I > B10 Asp > insulin-like growth factor-II > insulin > B28 Asp > B9 Asp/B27 Glu > proinsulin. The maximal effect of insulin-like growth factor-I on 3H-thymidine incorporation was 71±16% higher than the maximal effect of insulin. The maximal effect of insulin-like growth factor-II was at least as high as the effect of insulin-like growth factor-I. Furthermore, the maximal effect of B10 Asp was 62±10% higher than the maximal effect of insulin. Insulin-like growth factor-I and B10 Asp tended to increase cell number more than insulin. In conclusion, this study shows that insulin analogues interact with different potencies with receptors for insulin and insulin-like growth factor-I in vascular smooth muscle cells and that insulin-like growth factors and the insulin analogue B10 Asp have more pronounced growth effects than insulin. Substitution of the amino acid Asp for His at position B10 in insulin makes the molecule more similar to insulin-like growth factor-I, chemically and probably also biologically.

Key words

Insulin insulin analogues insulin-like growth factors proliferation vascular smooth muscle cells 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Brange J, Ribel U, Hansen JF, Dodson G, Hansen MT, Havelund S, Melberg SG, Norris F, Norris K, Snel L, Sørensen AR, Voigt HO (1988) Monomeric insulins obtained by protein engineering and their medical implications. Nature 333: 679–682Google Scholar
  2. 2.
    Ullrich A, Bell JR, Chen EY, Herrera R, Petruzzelli LM, Dull TJ, Gray A, Coussens L, Liao Y-C, Tsubokawa M, Mason A, Seeburg PH, Grundfeld C, Rosen OM, Ramachandran J (1985) Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature 313: 756–761Google Scholar
  3. 3.
    Ullrich A, Gray A, Tam AW, Yang-Feng T, Tsubokawa M, Collins C, Henzel W, Le Bon T, Kathuria S, Chen E, Jacobs S, Francke U, Ramachandran J, Fujita-Yamaguchi Y (1986) Insulin-like growth factor I receptor primary structure: comparison with insulin receptor suggests structural determinants that define functional specificity. EMBO J 5: 2503–2512Google Scholar
  4. 4.
    King GL, Goodman AD, Buzney S, Moses A, Kahn CR (1985) Receptors and growth-promoting effects of insulin and insulin- like growth factors on cells from bovine retinal capillaries and aorta. J Clin Invest 75: 1028–1036Google Scholar
  5. 5.
    Bornfeldt KE, Arnqvist HJ, Dahlkvist HH, Skottner A, Wikberg JES (1988) Receptors for insulin-like growth factor-I in plasma membranes isolated from bovine mesenteric arteries. Acta Endocrinol 117: 428–434Google Scholar
  6. 6.
    Pfeifle B, Ditschuneit HH, Ditschuneit H (1982) Binding and biological actions of insulin-like growth factors on human arterial smooth muscle cells. Horm Metab Res 14: 409–414Google Scholar
  7. 7.
    Kaiser N, Tur-Sinai A, Hasin M, Cerasi E (1985) Binding, degradation, and biological activity of insulin in vascular smooth muscle cells. Am J Physiol 249: E292-E298Google Scholar
  8. 8.
    Cascieri MA, Chicchi GG, Hayes NS, Slater EE (1986) Stimulation of DNA synthesis in rat A10 vascular smooth muscle cells by threonine-59 insulin-like growth factor-I. Circ Res 59: 171–177Google Scholar
  9. 9.
    Lee PDK, Hintz RL, Rosenfeld RG, Benitz WE (1988) Presence of insulin-like growth factor receptors and lack of insulin receptors on fetal bovine smooth muscle cells. In Vitro Cell Devel Biol 24: 921–926Google Scholar
  10. 10.
    Stout RW (1987) Insulin and atheroma — an update. Lancet I: 1077–1079Google Scholar
  11. 11.
    King GL (1985) Cell biology as an approach to the study of the vascular complications of diabetes. Metabolism 34 [Suppl. 1]: 17–24Google Scholar
  12. 12.
    Nilsson J, Ksiazek T, Heldin C-H, Thyberg J (1983) Demonstration of stimulatory effects of platelet-derived growth factor on cultivated rat arterial smooth muscle cells. Exp Cell Res 145: 231–237Google Scholar
  13. 13.
    Ham RG (1965) Clonal growth of mammalian cells in a chemically defined, synthetic medium. Proc Natl Acad Sci USA 53: 288–293Google Scholar
  14. 14.
    Ross R (1971) The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol 50: 172–186Google Scholar
  15. 15.
    Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillssen D, Gabbiani G (1986) A monoclonal antibody against α-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103: 2787–2796Google Scholar
  16. 16.
    Nilsson J, Thyberg J (1982) Fine structure of arterial smooth muscle cells cultured in the presence of whole blood serum or plasma-derived serum. Cell Tissue Res 223: 87–99Google Scholar
  17. 17.
    Nilsson J, Ksiazek T, Thyberg J (1983) Effects of colchicine on DNA synthesis, endocytosis and fine structure of cultivated arterial smooth muscle cells. Exp Cell Res 143: 367–375Google Scholar
  18. 18.
    Goldstein JL, Brown MS (1974) Binding and degradation of lowdensity lipoproteins by cultured human fibroblasts. J Biol Chem 249: 5153–5162Google Scholar
  19. 19.
    Skottner A, Fryklund L, Hansson HA (1986) Experimental research on IGF-I. Acta Paediatr Scand 325: 107–111Google Scholar
  20. 20.
    Waud DR (1975) Analysis of dose-response curves. In: Daniel EE, Paton DM (eds) Methods in pharmacology, Vol 3. Plenum Press, New York, pp 471–506Google Scholar
  21. 21.
    Marquardt DW (1963) An algorithm for least-squares estimation of non-linear parameters. J Soc Indust Appl Math 11: 431–441Google Scholar
  22. 22.
    Drejer K, Kruse V, Larsen UD (1988) Insulin analogs: binding to the human liver cell line, HEP G2. Diabetes Res Clin Pract 5 [Suppl 1]: 231Google Scholar
  23. 23.
    Limbird E (1986) tCell surface receptors: a short course on theory and methods, 3rd edn. Martinus Nijhoff, BostonGoogle Scholar
  24. 24.
    Elgin RG, Busby WH Jr, Clemmons DR (1987) An insulin-like growth factor (IGF) binding protein enhances the biologic response to IGF-I. Proc Natl Acad Sci USA 84: 3254–3258Google Scholar
  25. 25.
    Ross M, Francis GL, Szabo L, Wallace JC, Ballard FJ (1989) Insulin-like growth factor (IGF)-binding proteins inhibit the biological activities of IGF-1 and IGF-2 but not des-(1–3)-IGF-1. Biochem J 258: 267–272Google Scholar
  26. 26.
    Ritvos O, Ranta T, Jalkanen J, Suikkari A-M, Voutilainen R, Bohn H, Rutanen E-M (1988) Insulin-like growth factor (IGF) binding protein from human decidua inhibits the binding and biological action of IGF-I in cultured choriocarcinoma cells. Endocrinology 122: 2150–2157Google Scholar
  27. 27.
    McCusker RH, Clemmons DR (1988) Insulin-like growth factor binding protein secretion by muscle cells: effect of cellular differentiation and proliferation. J Cell Physiol 137: 505–512Google Scholar
  28. 28.
    Clemmons DR, Elgin RG, Han VKM, Casella SJ, D'Ercole AJ, Van Wyk JJ (1986) Cultured fibroblast monolayers secrete aprotein that alters the cellular binding of somatomedin-C/insulinlike growth factor I. J Clin Invest 77: 1548–1556Google Scholar
  29. 29.
    Bowman WC, Rand MJ (1980) Principles of drug action. In: Textbook of pharmacology, 2nd English edn. Blackwell Scientific Publications, Oxford, pp 1.39–69Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • K. E. Bornfeldt
    • 1
  • R. A. Gidlöf
    • 1
  • A. Wasteson
    • 2
  • M. Lake
    • 4
  • A. Skottner
    • 5
  • H. J. Arnqvist
    • 3
  1. 1.Department of Pharmacology, Faculty of Health SciencesLinköping UniversityLinköping
  2. 2.Department of Cell Biology, Faculty of Health SciencesLinköping UniversityLinköping
  3. 3.Department of Internal Medicine, Faculty of Health SciencesLinköping UniversityLinköping
  4. 4.Department of KabiGenStockholmSweden
  5. 5.Department of Kabi Peptide HormonesStockholmSweden

Personalised recommendations