Platelet-derived growth factor (PDGF) in oncogenesis: development of a vascular connective tissue stroma in xenotransplanted human melanoma producing PDGF-BB (original) (raw)

Abstract

Human WM9 melanoma cells, previously shown to be devoid of PDGF expression, were stably transfected with a PDGF-B cDNA under the transcriptional control of a cytomegalovirus promoter. Northern blot analysis revealed high expression of an mRNA of the expected size in the PDGF-B-transfected cells. Synthesis and secretion of PDGF-BB was confirmed by immunoprecipitation. Furthermore, conditioned medium from PDGF-B-transfected cells contained a mitogenic activity for fibroblasts. For analysis of tumor growth in vivo, cells of each type were injected subcutaneously into BALB/c nu/nu mice. Tumors from mice injected with WM9 cells transfected with the vector only contained large necrotic areas; only scant blood vessels with narrow lumina were observed. No connective tissue was present. In the tumors from PDGF-B-transfected WM9 cells, nests of tumor were divided by connective tissue septa. An abundance of blood vessels was observed in the connective tissue septa and within the tumor cell nests. There was a complete absence of necrosis in these tumors. The present results suggest that tumor-derived PDGF-BB is a potent mediator of connective tissue stroma formation. The connective tissue framework that is generated in response to PDGF-BB may form a solid support for newly formed blood vessels and, thereby, facilitate the formation of a functional vascular system in the tumor.

393

Images in this article

Selected References

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

  1. 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]
  2. Beitz J. G., Kim I. S., Calabresi P., Frackelton A. R., Jr Human microvascular endothelial cells express receptors for platelet-derived growth factor. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):2021–2025. doi: 10.1073/pnas.88.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Betsholtz C., Westermark B. Growth factor-induced proliferation of human fibroblasts in serum-free culture depends on cell density and extracellular calcium concentration. J Cell Physiol. 1984 Feb;118(2):203–210. doi: 10.1002/jcp.1041180213. [DOI] [PubMed] [Google Scholar]
  4. Bonthron D. T., Sultan P., Collins T. Structure of the murine c-sis proto-oncogene (Sis, PDGFB) encoding the B chain of platelet-derived growth factor. Genomics. 1991 May;10(1):287–292. doi: 10.1016/0888-7543(91)90515-g. [DOI] [PubMed] [Google Scholar]
  5. Bywater M., Rorsman F., Bongcam-Rudloff E., Mark G., Hammacher A., Heldin C. H., Westermark B., Betsholtz C. Expression of recombinant platelet-derived growth factor A- and B-chain homodimers in rat-1 cells and human fibroblasts reveals differences in protein processing and autocrine effects. Mol Cell Biol. 1988 Jul;8(7):2753–2762. doi: 10.1128/mcb.8.7.2753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deuel T. F., Senior R. M., Huang J. S., Griffin G. L. Chemotaxis of monocytes and neutrophils to platelet-derived growth factor. J Clin Invest. 1982 Apr;69(4):1046–1049. doi: 10.1172/JCI110509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Devare S. G., Reddy E. P., Law J. D., Robbins K. C., Aaronson S. A. Nucleotide sequence of the simian sarcoma virus genome: demonstration that its acquired cellular sequences encode the transforming gene product p28sis. Proc Natl Acad Sci U S A. 1983 Feb;80(3):731–735. doi: 10.1073/pnas.80.3.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Doolittle R. F., Hunkapiller M. W., Hood L. E., Devare S. G., Robbins K. C., Aaronson S. A., Antoniades H. N. Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor. Science. 1983 Jul 15;221(4607):275–277. doi: 10.1126/science.6304883. [DOI] [PubMed] [Google Scholar]
  9. Dvorak H. F. Tumors: wounds that do not heal. Similarities between tumor stroma generation and wound healing. N Engl J Med. 1986 Dec 25;315(26):1650–1659. doi: 10.1056/NEJM198612253152606. [DOI] [PubMed] [Google Scholar]
  10. Eva A., Robbins K. C., Andersen P. R., Srinivasan A., Tronick S. R., Reddy E. P., Ellmore N. W., Galen A. T., Lautenberger J. A., Papas T. S. Cellular genes analogous to retroviral onc genes are transcribed in human tumour cells. Nature. 1982 Jan 14;295(5845):116–119. doi: 10.1038/295116a0. [DOI] [PubMed] [Google Scholar]
  11. Folkman J. How is blood vessel growth regulated in normal and neoplastic tissue? G.H.A. Clowes memorial Award lecture. Cancer Res. 1986 Feb;46(2):467–473. [PubMed] [Google Scholar]
  12. Folkman J. What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst. 1990 Jan 3;82(1):4–6. doi: 10.1093/jnci/82.1.4. [DOI] [PubMed] [Google Scholar]
  13. Hammacher A., Nistér M., Westermark B., Heldin C. H. A human glioma cell line secretes three structurally and functionally different dimeric forms of platelet-derived growth factor. Eur J Biochem. 1988 Sep 1;176(1):179–186. doi: 10.1111/j.1432-1033.1988.tb14266.x. [DOI] [PubMed] [Google Scholar]
  14. Hart C. E., Forstrom J. W., Kelly J. D., Seifert R. A., Smith R. A., Ross R., Murray M. J., Bowen-Pope D. F. Two classes of PDGF receptor recognize different isoforms of PDGF. Science. 1988 Jun 10;240(4858):1529–1531. doi: 10.1126/science.2836952. [DOI] [PubMed] [Google Scholar]
  15. Heldin C. H., Bäckström G., Ostman A., Hammacher A., Rönnstrand L., Rubin K., Nistér M., Westermark B. Binding of different dimeric forms of PDGF to human fibroblasts: evidence for two separate receptor types. EMBO J. 1988 May;7(5):1387–1393. doi: 10.1002/j.1460-2075.1988.tb02955.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heldin C. H., Westermark B. Platelet-derived growth factor: mechanism of action and possible in vivo function. Cell Regul. 1990 Jul;1(8):555–566. doi: 10.1091/mbc.1.8.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Heldin N. E., Cvejić D., Smeds S., Westermark B. Coexpression of functionally active receptors for thyrotropin and platelet-derived growth factor in human thyroid carcinoma cells. Endocrinology. 1991 Oct;129(4):2187–2193. doi: 10.1210/endo-129-4-2187. [DOI] [PubMed] [Google Scholar]
  19. Herlyn M., Balaban G., Bennicelli J., Guerry D., 4th, Halaban R., Herlyn D., Elder D. E., Maul G. G., Steplewski Z., Nowell P. C. Primary melanoma cells of the vertical growth phase: similarities to metastatic cells. J Natl Cancer Inst. 1985 Feb;74(2):283–289. [PubMed] [Google Scholar]
  20. Hermansson M., Nistér M., Betsholtz C., Heldin C. H., Westermark B., Funa K. Endothelial cell hyperplasia in human glioblastoma: coexpression of mRNA for platelet-derived growth factor (PDGF) B chain and PDGF receptor suggests autocrine growth stimulation. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7748–7752. doi: 10.1073/pnas.85.20.7748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jain R. K. Transport of molecules in the tumor interstitium: a review. Cancer Res. 1987 Jun 15;47(12):3039–3051. [PubMed] [Google Scholar]
  22. Khachigian L. M., Owensby D. A., Chesterman C. N. A tyrosinated peptide representing the alternatively spliced exon of the platelet-derived growth factor A-chain binds specifically to cultured cells and interferes with binding of several growth factors. J Biol Chem. 1992 Jan 25;267(3):1660–1666. [PubMed] [Google Scholar]
  23. LaRochelle W. J., May-Siroff M., Robbins K. C., Aaronson S. A. A novel mechanism regulating growth factor association with the cell surface: identification of a PDGF retention domain. Genes Dev. 1991 Jul;5(7):1191–1199. doi: 10.1101/gad.5.7.1191. [DOI] [PubMed] [Google Scholar]
  24. Leveen P., Claesson-Welsh L., Heldin C. H., Westermark B., Betsholtz C. Expression of messenger RNAs for platelet-derived growth factor and its receptors in human sarcoma cell lines. Int J Cancer. 1990 Dec 15;46(6):1066–1070. doi: 10.1002/ijc.2910460620. [DOI] [PubMed] [Google Scholar]
  25. Nistér M., Claesson-Welsh L., Eriksson A., Heldin C. H., Westermark B. Differential expression of platelet-derived growth factor receptors in human malignant glioma cell lines. J Biol Chem. 1991 Sep 5;266(25):16755–16763. [PubMed] [Google Scholar]
  26. Ostman A., Andersson M., Betsholtz C., Westermark B., Heldin C. H. Identification of a cell retention signal in the B-chain of platelet-derived growth factor and in the long splice version of the A-chain. Cell Regul. 1991 Jul;2(7):503–512. doi: 10.1091/mbc.2.7.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ostman A., Rall L., Hammacher A., Wormstead M. A., Coit D., Valenzuela P., Betsholtz C., Westermark B., Heldin C. H. Synthesis and assembly of a functionally active recombinant platelet-derived growth factor AB heterodimer. J Biol Chem. 1988 Nov 5;263(31):16202–16208. [PubMed] [Google Scholar]
  28. Pech M., Gazit A., Arnstein P., Aaronson S. A. Generation of fibrosarcomas in vivo by a retrovirus that expresses the normal B chain of platelet-derived growth factor and mimics the alternative splice pattern of the v-sis oncogene. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2693–2697. doi: 10.1073/pnas.86.8.2693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Powe J., Pak K. Y., Paik C. H., Steplewski Z., Ebbert M. A., Herlyn D., Ernst C., Alavi A., Eckelman W. C., Reba R. C. Labeling monoclonal antibodies and F(ab')2 fragments with (111In) indium using cyclic DTPA anhydride and their in vivo behavior in mice bearing human tumor xenografts. Cancer Drug Deliv. 1984;1(2):125–135. doi: 10.1089/cdd.1984.1.125. [DOI] [PubMed] [Google Scholar]
  30. Raines E. W., Ross R. Compartmentalization of PDGF on extracellular binding sites dependent on exon-6-encoded sequences. J Cell Biol. 1992 Jan;116(2):533–543. doi: 10.1083/jcb.116.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rao C. D., Igarashi H., Chiu I. M., Robbins K. C., Aaronson S. A. Structure and sequence of the human c-sis/platelet-derived growth factor 2 (SIS/PDGF2) transcriptional unit. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2392–2396. doi: 10.1073/pnas.83.8.2392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rao C. D., Pech M., Robbins K. C., Aaronson S. A. The 5' untranslated sequence of the c-sis/platelet-derived growth factor 2 transcript is a potent translational inhibitor. Mol Cell Biol. 1988 Jan;8(1):284–292. doi: 10.1128/mcb.8.1.284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Robbins K. C., Antoniades H. N., Devare S. G., Hunkapiller M. W., Aaronson S. A. Structural and immunological similarities between simian sarcoma virus gene product(s) and human platelet-derived growth factor. Nature. 1983 Oct 13;305(5935):605–608. doi: 10.1038/305605a0. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Seppä H., Grotendorst G., Seppä S., Schiffmann E., Martin G. R. Platelet-derived growth factor in chemotactic for fibroblasts. J Cell Biol. 1982 Feb;92(2):584–588. doi: 10.1083/jcb.92.2.584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  38. Siegbahn A., Hammacher A., Westermark B., Heldin C. H. Differential effects of the various isoforms of platelet-derived growth factor on chemotaxis of fibroblasts, monocytes, and granulocytes. J Clin Invest. 1990 Mar;85(3):916–920. doi: 10.1172/JCI114519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Smits A., Hermansson M., Nistér M., Karnushina I., Heldin C. H., Westermark B., Funa K. Rat brain capillary endothelial cells express functional PDGF B-type receptors. Growth Factors. 1989;2(1):1–8. doi: 10.3109/08977198909069076. [DOI] [PubMed] [Google Scholar]
  40. Thyberg J., Ostman A., Bäckström G., Westermark B., Heldin C. H. Localization of platelet-derived growth factor (PDGF) in CHO cells transfected with PDGF A- or B-chain cDNA: retention of PDGF-BB in the endoplasmic reticulum and Golgi complex. J Cell Sci. 1990 Oct;97(Pt 2):219–229. doi: 10.1242/jcs.97.2.219. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. Westermark B., Heldin C. H. Platelet-derived growth factor in autocrine transformation. Cancer Res. 1991 Oct 1;51(19):5087–5092. [PubMed] [Google Scholar]
  43. Westermark B., Johnsson A., Paulsson Y., Betsholtz C., Heldin C. H., Herlyn M., Rodeck U., Koprowski H. Human melanoma cell lines of primary and metastatic origin express the genes encoding the chains of platelet-derived growth factor (PDGF) and produce a PDGF-like growth factor. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7197–7200. doi: 10.1073/pnas.83.19.7197. [DOI] [PMC free article] [PubMed] [Google Scholar]