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. 1988 Feb 1;106(2):403–413. doi: 10.1083/jcb.106.2.403

Arterial smooth muscle cells express platelet-derived growth factor (PDGF) A chain mRNA, secrete a PDGF-like mitogen, and bind exogenous PDGF in a phenotype- and growth state-dependent manner

PMCID: PMC2114975  PMID: 2828383

Abstract

Adult rat arterial smooth muscle cells are shown to express platelet- derived growth factor (PDGF) A chain mRNA, to secrete a PDGF-like mitogen, and to bind exogenous PDGF in a phenotype- and growth state- dependent manner. In the intact aortic media, where the cells are in a contractile phenotype, only minute amounts of PDGF A chain and no B chain (c-sis) RNA were detected. After cultivation and modulation of the cells into a synthetic phenotype, the A chain gene was distinctly expressed, whereas the B chain gene remained unexpressed. Cells kept in serum-free medium on a substrate of plasma fibronectin showed high levels of A chain RNA and high PDGF receptor activity, but did not secrete detectable amounts of PDGF-like mitogen. After exposure to PDGF, which is itself sufficient to initiate DNA synthesis and mitosis in these cells, a PDGF-like mitogen was released into the extracellular medium. Concomitantly, the amount of A chain transcripts per cell and the ability of the cells to bind radioactive PDGF decreased. Similarly, smooth muscle cells initially grown in the presence of serum released more PDGF-like mitogen, contained fewer A chain transcripts, and bound more radioactive PDGF in proliferating than in stationary cultures. The findings confirm the notion that adult rat arterial smooth muscle cells are able to promote their own growth in an autocrine or paracrine manner. Furthermore, they reveal some basic principles in the control of this process.

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Selected References

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  1. Assoian R. K., Sporn M. B. Type beta transforming growth factor in human platelets: release during platelet degranulation and action on vascular smooth muscle cells. J Cell Biol. 1986 Apr;102(4):1217–1223. doi: 10.1083/jcb.102.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  3. Benya P. D., Shaffer J. D. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 1982 Aug;30(1):215–224. doi: 10.1016/0092-8674(82)90027-7. [DOI] [PubMed] [Google Scholar]
  4. Betsholtz C., Johnsson A., Heldin C. H., Westermark B. Efficient reversion of simian sarcoma virus-transformation and inhibition of growth factor-induced mitogenesis by suramin. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6440–6444. doi: 10.1073/pnas.83.17.6440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Betsholtz C., Johnsson A., Heldin C. H., Westermark B., Lind P., Urdea M. S., Eddy R., Shows T. B., Philpott K., Mellor A. L. cDNA sequence and chromosomal localization of human platelet-derived growth factor A-chain and its expression in tumour cell lines. Nature. 1986 Apr 24;320(6064):695–699. doi: 10.1038/320695a0. [DOI] [PubMed] [Google Scholar]
  6. Burke J. M., Ross R. Synthesis of connective tissue macromolecules by smooth muscle. Int Rev Connect Tissue Res. 1979;8:119–157. doi: 10.1016/b978-0-12-363708-6.50010-2. [DOI] [PubMed] [Google Scholar]
  7. Chamley-Campbell J. H., Campbell G. R., Ross R. Phenotype-dependent response of cultured aortic smooth muscle to serum mitogens. J Cell Biol. 1981 May;89(2):379–383. doi: 10.1083/jcb.89.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chamley-Campbell J., Campbell G. R., Ross R. The smooth muscle cell in culture. Physiol Rev. 1979 Jan;59(1):1–61. doi: 10.1152/physrev.1979.59.1.1. [DOI] [PubMed] [Google Scholar]
  9. Chamley J. H., Campbell G. R., McConnell J. D., Gröschel-Stewart U. Comparison of vascular smooth muscle cells from adult human, monkey and rabbit in primary culture and in subculture. Cell Tissue Res. 1977 Feb 14;177(4):503–522. doi: 10.1007/BF00220611. [DOI] [PubMed] [Google Scholar]
  10. Clarke M. F., Westin E., Schmidt D., Josephs S. F., Ratner L., Wong-Staal F., Gallo R. C., Reitz M. S., Jr Transformation of NIH 3T3 cells by a human c-sis cDNA clone. 1984 Mar 29-Apr 4Nature. 308(5958):464–467. doi: 10.1038/308464a0. [DOI] [PubMed] [Google Scholar]
  11. Edmonds M., Caramela M. G. The isolation and characterization of adenosine monophosphate-rich polynucleotides synthesized by Ehrlich ascites cells. J Biol Chem. 1969 Mar 10;244(5):1314–1324. [PubMed] [Google Scholar]
  12. Fritz K. E., Jarmolych J., Daoud A. S. Association of DNA synthesis and apparent dedifferentiation of aortic smooth muscle cells in vitro. Exp Mol Pathol. 1970 Jun;12(3):354–362. doi: 10.1016/0014-4800(70)90066-3. [DOI] [PubMed] [Google Scholar]
  13. Gabbiani G., Kocher O., Bloom W. S., Vandekerckhove J., Weber K. Actin expression in smooth muscle cells of rat aortic intimal thickening, human atheromatous plaque, and cultured rat aortic media. J Clin Invest. 1984 Jan;73(1):148–152. doi: 10.1172/JCI111185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heldin C. H., Johnsson A., Ek B., Wennergren S., Rönnstrand L., Hammacher A., Faulders B., Wasteson A., Westermark B. Purification of human platelet-derived growth factor. Methods Enzymol. 1987;147:3–13. doi: 10.1016/0076-6879(87)47094-8. [DOI] [PubMed] [Google Scholar]
  15. Heldin C. H., Wasteson A., Westermark B. Interaction of platelet-derived growth factor with its fibroblast receptor. Demonstration of ligand degradation and receptor modulation. J Biol Chem. 1982 Apr 25;257(8):4216–4221. [PubMed] [Google Scholar]
  16. Heldin C. H., Wasteson A., Westermark B. Platelet-derived growth factor. Mol Cell Endocrinol. 1985 Mar;39(3):169–187. doi: 10.1016/0303-7207(85)90061-9. [DOI] [PubMed] [Google Scholar]
  17. Heldin C. H., Westermark B. Growth factors: mechanism of action and relation to oncogenes. Cell. 1984 May;37(1):9–20. doi: 10.1016/0092-8674(84)90296-4. [DOI] [PubMed] [Google Scholar]
  18. Heldin C. H., Westermark B., Wasteson A. Demonstration of an antibody against platelet-derived growth factor. Exp Cell Res. 1981 Dec;136(2):255–261. doi: 10.1016/0014-4827(81)90003-3. [DOI] [PubMed] [Google Scholar]
  19. Heldin C. H., Westermark B., Wasteson A. Specific receptors for platelet-derived growth factor on cells derived from connective tissue and glia. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3664–3668. doi: 10.1073/pnas.78.6.3664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hosang M. Suramin binds to platelet-derived growth factor and inhibits its biological activity. J Cell Biochem. 1985;29(3):265–273. doi: 10.1002/jcb.240290310. [DOI] [PubMed] [Google Scholar]
  21. Johnsson A., Betsholtz C., Heldin C. H., Westermark B. The phenotypic characteristics of simian sarcoma virus-transformed human fibroblasts suggest that the v-sis gene product acts solely as a PDGF receptor agonist in cell transformation. EMBO J. 1986 Jul;5(7):1535–1541. doi: 10.1002/j.1460-2075.1986.tb04394.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Larson D. M., Fujiwara K., Alexander R. W., Gimbrone M. A., Jr Myosin in cultured vascular smooth muscle cells: immunofluorescence and immunochemical studies of alterations in antigenic expression. J Cell Biol. 1984 Nov;99(5):1582–1589. doi: 10.1083/jcb.99.5.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  24. Nilsson J., Sjölund M., Palmberg L., Thyberg J., Heldin C. H. Arterial smooth muscle cells in primary culture produce a platelet-derived growth factor-like protein. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4418–4422. doi: 10.1073/pnas.82.13.4418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nilsson J., Thyberg J., Heldin C. H., Westermark B., Wasteson A. Surface binding and internalization of platelet-derived growth factor in human fibroblasts. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5592–5596. doi: 10.1073/pnas.80.18.5592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nistér M., Heldin C. H., Wasteson A., Westermark B. A glioma-derived analog to platelet-derived growth factor: demonstration of receptor competing activity and immunological crossreactivity. Proc Natl Acad Sci U S A. 1984 Feb;81(3):926–930. doi: 10.1073/pnas.81.3.926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Owens G. K., Loeb A., Gordon D., Thompson M. M. Expression of smooth muscle-specific alpha-isoactin in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation. J Cell Biol. 1986 Feb;102(2):343–352. doi: 10.1083/jcb.102.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Palmberg L., Sjölund M., Thyberg J. Phenotype modulation in primary cultures of arterial smooth-muscle cells: reorganization of the cytoskeleton and activation of synthetic activities. Differentiation. 1985;29(3):275–283. doi: 10.1111/j.1432-0436.1985.tb00327.x. [DOI] [PubMed] [Google Scholar]
  29. Rosenfeld M. E., Bowen-Pope D. F., Ross R. Platelet-derived growth factor: morphologic and biochemical studies of binding, internalization, and degradation. J Cell Physiol. 1984 Nov;121(2):263–274. doi: 10.1002/jcp.1041210202. [DOI] [PubMed] [Google Scholar]
  30. Ross R., Raines E. W., Bowen-Pope D. F. The biology of platelet-derived growth factor. Cell. 1986 Jul 18;46(2):155–169. doi: 10.1016/0092-8674(86)90733-6. [DOI] [PubMed] [Google Scholar]
  31. Ross R. The pathogenesis of atherosclerosis--an update. N Engl J Med. 1986 Feb 20;314(8):488–500. doi: 10.1056/NEJM198602203140806. [DOI] [PubMed] [Google Scholar]
  32. Ruoslahti E., Hayman E. G., Pierschbacher M., Engvall E. Fibronectin: purification, immunochemical properties, and biological activities. Methods Enzymol. 1982;82(Pt A):803–831. doi: 10.1016/0076-6879(82)82103-4. [DOI] [PubMed] [Google Scholar]
  33. Schwartz S. M., Campbell G. R., Campbell J. H. Replication of smooth muscle cells in vascular disease. Circ Res. 1986 Apr;58(4):427–444. doi: 10.1161/01.res.58.4.427. [DOI] [PubMed] [Google Scholar]
  34. Seifert R. A., Schwartz S. M., Bowen-Pope D. F. Developmentally regulated production of platelet-derived growth factor-like molecules. Nature. 1984 Oct 18;311(5987):669–671. doi: 10.1038/311669a0. [DOI] [PubMed] [Google Scholar]
  35. Sejersen T., Betsholtz C., Sjölund M., Heldin C. H., Westermark B., Thyberg J. Rat skeletal myoblasts and arterial smooth muscle cells express the gene for the A chain but not the gene for the B chain (c-sis) of platelet-derived growth factor (PDGF) and produce a PDGF-like protein. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6844–6848. doi: 10.1073/pnas.83.18.6844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sejersen T., Sümegi J., Ringertz N. R. Density-dependent arrest of DNA replication is accompanied by decreased levels of c-myc mRNA in myogenic but not in differentiation-defective myoblasts. J Cell Physiol. 1985 Dec;125(3):465–470. doi: 10.1002/jcp.1041250315. [DOI] [PubMed] [Google Scholar]
  37. Shin S. I., Freedman V. H., Risser R., Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. doi: 10.1073/pnas.72.11.4435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sjölund M., Madsen K., von der Mark K., Thyberg J. Phenotype modulation in primary cultures of smooth-muscle cells from rat aorta. Synthesis of collagen and elastin. Differentiation. 1986;32(2):173–180. doi: 10.1111/j.1432-0436.1986.tb00570.x. [DOI] [PubMed] [Google Scholar]
  39. Skalli O., Bloom W. S., Ropraz P., Azzarone B., Gabbiani G. Cytoskeletal remodeling of rat aortic smooth muscle cells in vitro: relationships to culture conditions and analogies to in vivo situations. J Submicrosc Cytol. 1986 Jul;18(3):481–493. [PubMed] [Google Scholar]
  40. Sporn M. B., Todaro G. J. Autocrine secretion and malignant transformation of cells. N Engl J Med. 1980 Oct 9;303(15):878–880. doi: 10.1056/NEJM198010093031511. [DOI] [PubMed] [Google Scholar]
  41. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Thyberg J., Palmberg L., Nilsson J., Ksiazek T., Sjölund M. Phenotype modulation in primary cultures of arterial smooth muscle cells. On the role of platelet-derived growth factor. Differentiation. 1983;25(2):156–167. doi: 10.1111/j.1432-0436.1984.tb01351.x. [DOI] [PubMed] [Google Scholar]
  43. Traganos F., Darzynkiewicz Z., Sharpless T., Melamed M. R. Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system. J Histochem Cytochem. 1977 Jan;25(1):46–56. doi: 10.1177/25.1.64567. [DOI] [PubMed] [Google Scholar]
  44. Travo P., Weber K., Osborn M. Co-existence of vimentin and desmin type intermediate filaments in a subpopulation of adult rat vascular smooth muscle cells growing in primary culture. Exp Cell Res. 1982 May;139(1):87–94. doi: 10.1016/0014-4827(82)90321-4. [DOI] [PubMed] [Google Scholar]
  45. Walker L. N., Bowen-Pope D. F., Ross R., Reidy M. A. Production of platelet-derived growth factor-like molecules by cultured arterial smooth muscle cells accompanies proliferation after arterial injury. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7311–7315. doi: 10.1073/pnas.83.19.7311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Waterfield M. D., Scrace G. T., Whittle N., Stroobant P., Johnsson A., Wasteson A., Westermark B., Heldin C. H., Huang J. S., Deuel T. F. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature. 1983 Jul 7;304(5921):35–39. doi: 10.1038/304035a0. [DOI] [PubMed] [Google Scholar]
  47. Williams L. T., Tremble P. M., Lavin M. F., Sunday M. E. Platelet-derived growth factor receptors form a high affinity state in membrane preparations. Kinetics and affinity cross-linking studies. J Biol Chem. 1984 Apr 25;259(8):5287–5294. [PubMed] [Google Scholar]

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