Journal of Cancer Research and Clinical Oncology

, Volume 132, Issue 9, pp 589–599 | Cite as

Platelet-derived growth factor receptor independent proliferation of human glioblastoma cells: selective tyrosine kinase inhibitors lack antiproliferative activity

  • Dietmar Gross
  • Günther Bernhardt
  • Armin BuschauerEmail author
Original Paper


Purpose: The aim of this study was to investigate the role of platelet-derived growth factor (PDGF) and PDGF receptors (PDGFRs) in the proliferation of human glioblastoma cells as a prerequisite for a new therapeutic approach to the treatment of malignant brain tumors with selective tyrosine kinase inhibitors such as imatinib. Methods and results: In the human glioblastoma cell lines U-87 MG, U-118 MG and U-373 MG different PDGF and PDGFR mRNAs were detected by RT-PCR, and the expression of the receptor proteins was demonstrated by immunostaining and flow cytometry. Moreover, functional activity of PDGFRs was demonstrated in PDGFRβ expressing glioblastoma cell variants by measuring the mobilization of intracellular Ca2+ upon PDGF-BB stimulation. However, addition of PDGF-BB to the serum-free culture medium had no stimulatory effect on cell proliferation. Furthermore, cell growth in serum-supplemented and serum-free medium was not affected by imatinib, leflunomide and AG-1296 at therapeutically relevant concentrations. Conclusion: Our results suggest that clinical antitumor effects of imatinib on glioblastoma, if any, are not mediated by the PDGFR.


Platelet-derived growth factor receptor Human glioblastoma cells Imatinib 



Chronic myelogenous leukemia


Fetal calf serum


Dimethyl sulfoxide


Fluorescein isothiocyanate


3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide


Platelet-derived growth factor


Platelet-derived growth factor receptor


Recombinant human PDGF-BB


Reverse transcription polymerase chain reaction


Vascular endothelial growth factor


Basic fibroblast growth factor


  1. Bernhardt G, Reile H, Birnböck H, Spruss T, Schönenberger H (1992) Standardized kinetic microassay to quantify differential chemosensitivity on the basis of proliferative activity. J Cancer Res Clin Oncol 118(1):35–43PubMedCrossRefGoogle Scholar
  2. Berridge MJ, Heslop JP, Irvine RF, Brown KD (1984) Inositol trisphosphate formation and calcium mobilization in Swiss 3T3 cells in response to platelet-derived growth factor. Biochem J 222(1):195–201PubMedGoogle Scholar
  3. Böhmer FD, Karagyozov L, Uecker A, Serve H, Botzki A, Mahboobi S, Dove S (2003) A single amino acid exchange inverts susceptibility of related receptor tyrosine kinases for the ATP site inhibitor STI-571. J Biol Chem 278(7):5148–5155PubMedCrossRefGoogle Scholar
  4. Buchdunger E, O’Reilly T, Wood J (2002) Pharmacology of imatinib (STI571). Eur J Cancer 38(Suppl. 5):28–36CrossRefGoogle Scholar
  5. Cherrington JM, Strawn LM, Shawver LK (2000) New paradigms for the treatment of cancer: the role of anti-angiogenesis agents. Adv Cancer Res 79:1–38PubMedGoogle Scholar
  6. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 162(1):156–159PubMedCrossRefGoogle Scholar
  7. Denk PO, Knorr M (2002) Differential regulation of expression of PDGF receptors on corneal epithelial cells. Ophthalmologe 99(1):15–19PubMedCrossRefGoogle Scholar
  8. Diliberto PA, Gordon GW, Yu CL, Earp HS, Herman B (1992) Platelet-derived growth factor (PDGF) alpha receptor activation modulates the calcium mobilizing activity of the PDGF beta receptor in Balb/c3T3 fibroblasts. J Biol Chem 267(17):11888–11897PubMedGoogle Scholar
  9. Dresemann G (2003) STI 571/hydroxyurea in progressive, pretreated glioblastoma (GB) patients (pts.). In: 39th ASCO annual meeting, Chicago, May 31–June 3 2003, abstract no. 465Google Scholar
  10. Dresemann G (2004) Imatinib (STI571) plus hydroxyurea: safety and efficacy in pretreated, progressive glioblastoma multiforme (GBM) patients (pts). In: 40th ASCO annual meeting, New Orleans, 5–8 June 2004, abstract no. 1550Google Scholar
  11. Eckhardt SG, Rizzo J, Sweeney KR, Cropp G, Baker SD, Kraynak MA, Kuhn JG, Villalona-Calero MA, Hammond L, Weiss G, Thurman A, Smith L, Drengler R, Eckardt JR, Moczygemba J, Hannah AL, Von Hoff DD, Rowinsky EK (1999) Phase I and pharmacologic study of the tyrosine kinase inhibitor SU101 in patients with advanced solid tumors. J Clin Oncol 17(4):1095–1104PubMedGoogle Scholar
  12. Gambacorti-Passerini C, le Coutre P, Mologni L, Fanelli M, Bertazzoli C, Marchesi E, Di Nicola M, Biondi A, Corneo GM, Belotti D, Pogliani E, Lydon NB (1997) Inhibition of the ABL kinase activity blocks the proliferation of BCR/ABL+ leukemic cells and induces apoptosis. Blood Cells Mol Dis 23(3):380–394PubMedCrossRefGoogle Scholar
  13. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260(6):3440–3450PubMedGoogle Scholar
  14. Guha A, Dashner K, Black PM, Wagner JA, Stiles CD (1995) Expression of PDGF and PDGF receptors in human astrocytoma operation specimens supports the existence of an autocrine loop. Int J Cancer 60(2):168–173PubMedGoogle Scholar
  15. Gupta M, Naik S, Pandey CM, Dabadghao S (2002) Drug sensitivity assay for leukaemic cells by flow cytometry. Indian J Med Res 115:260–264PubMedGoogle Scholar
  16. Hermanson M, Funa K, Hartman M, Claesson-Welsh L, Heldin CH, Westermark B, Nister M (1992) Platelet-derived growth factor and its receptors in human glioma tissue: expression of messenger RNA and protein suggests the presence of autocrine and paracrine loops. Cancer Res 52(11):3213–3219PubMedGoogle Scholar
  17. Holdhoff M, Kreuzer KA, Appelt C, Scholz R, Na IK, Hildebrandt B, Riess H, Jordan A, Schmidt CA, Van Etten RA, Dorken B, le Coutre P (2005) Imatinib mesylate radiosensitizes human glioblastoma cells through inhibition of platelet-derived growth factor receptor. Blood Cells Mol Dis 34(2):181–185PubMedCrossRefGoogle Scholar
  18. Kilic T, Alberta JA, Zdunek PR, Acar M, Iannarelli P, O’Reilly T, Buchdunger E, Black PM, Stiles CD (2000) Intracranial inhibition of platelet-derived growth factor-mediated glioblastoma cell growth by an orally active kinase inhibitor of the 2-phenylaminopyrimidine class. Cancer Res 60(18):5143–5150PubMedGoogle Scholar
  19. Kovalenko M, Gazit A, Böhmer A, Rorsman C, Ronnstrand L, Heldin CH, Waltenberger J, Böhmer FD, Levitzki A (1994) Selective platelet-derived growth factor receptor kinase blockers reverse sis-transformation. Cancer Res 54(23):6106–6114PubMedGoogle Scholar
  20. Lokker NA, Sullivan CM, Hollenbach SJ, Israel MA, Giese NA (2002) Platelet-derived growth factor (PDGF) autocrine signaling regulates survival and mitogenic pathways in glioblastoma cells: evidence that the novel PDGF-C and PDGF-D ligands may play a role in the development of brain tumors. Cancer Res 62(13):3729–3735PubMedGoogle Scholar
  21. Lyseng-Williamson K, Jarvis B (2001) Imatinib. Drugs 61(12):1765–1774; discussion 1775–1776Google Scholar
  22. McGary EC, Onn A, Mills L, Heimberger A, Eton O, Thomas GW, Shtivelband M, Bar-Eli M (2004) Imatinib mesylate inhibits platelet-derived growth factor receptor phosphorylation of melanoma cells but does not affect tumorigenicity in vivo. J Invest Dermatol 122(2):400–405PubMedCrossRefGoogle Scholar
  23. Moolenaar WH, Tertoolen LG, de Laat SW (1984) Growth factors immediately raise cytoplasmic free Ca2+ in human fibroblasts. J Biol Chem 259(13):8066–8069PubMedGoogle Scholar
  24. Morin MJ (2000) From oncogene to drug: development of small molecule tyrosine kinase inhibitors as anti-tumor and anti-angiogenic agents. Oncogene 19(56):6574–6583PubMedCrossRefGoogle Scholar
  25. National Cancer Institute (2005) Phase II study of imatinib mesylate in patients with gliomas.
  26. Nister M, Claesson-Welsh L, Eriksson A, Heldin CH, Westermark B (1991) Differential expression of platelet-derived growth factor receptors in human malignant glioma cell lines. J Biol Chem 266(25):16755–16763PubMedGoogle Scholar
  27. Ostman A, Böhmer FD (2001) Regulation of receptor tyrosine kinase signaling by protein tyrosine phosphatases. Trends Cell Biol 11(6):258–266PubMedCrossRefGoogle Scholar
  28. Ostman A, Heldin CH (2001) Involvement of platelet-derived growth factor in disease: development of specific antagonists. Adv Cancer Res 80:1–38PubMedCrossRefGoogle Scholar
  29. Pollack IF, Randall MS, Kristofik MP, Kelly RH, Selker RG, Vertosick FT (1990) Response of malignant glioma cell lines to epidermal growth factor and platelet-derived growth factor in a serum-free medium. J Neurosurg 73(1):106–112PubMedCrossRefGoogle Scholar
  30. Randal J (2000) Antiangiogenesis drugs target specific cancers, mechanisms. J Natl Cancer Inst 92(7):520–522PubMedCrossRefGoogle Scholar
  31. Raymond E, Brandes A, Van Oosterom A, Dittrich C, Fumoleau P, Coudert B, Twelves C, De Balincourt C, Lacombe D, Van Den Bent M (2004) Multicentre phase II study of imatinib mesylate in patients with recurrent glioblastoma: an EORTC: NDDG/BTG Intergroup study. J Clin Oncol 22(Suppl. 14):A-1501Google Scholar
  32. Sargent JM (2003) The use of the MTT assay to study drug resistance in fresh tumour samples. Recent Results Cancer Res 161:13–25PubMedGoogle Scholar
  33. Singer HS, Hansen B, Martinie D, Karp CL (1999) Mitogenesis in glioblastoma multiforme cell lines: a role for NGF and its TrkA receptors. J Neurooncol 45(1):1–8PubMedCrossRefGoogle Scholar
  34. Soria JC, Johnson BE, Chevalier TL (2003) Imatinib in small cell lung cancer. Lung Cancer 41(Suppl. 1):49–53CrossRefGoogle Scholar
  35. Steck PA, Gallick GE, Maxwell SA, Kloetzer WS, Arlinghaus RB, Moser RP, Gutterman JU, Yung WK (1986) Expression of epidermal growth factor receptor and associated glycoprotein on cultured human brain tumor cells. J Cell Biochem 32(1):1–10PubMedCrossRefGoogle Scholar
  36. Sultana S, Zhou R, Sadagopan MS, Skalli O (1998) Effects of growth factors and basement membrane proteins on the phenotype of U-373 MG glioblastoma cells as determined by the expression of intermediate filament proteins. Am J Pathol 153(4):1157–1168PubMedGoogle Scholar
  37. Takahashi A, Camacho P, Lechleiter JD, Herman B (1999) Measurement of intracellular calcium. Physiol Rev 79(4):1089–1125PubMedGoogle Scholar
  38. U.S. National Institutes of Health (2005a) Imatinib mesylate in treating patients with gliomas.
  39. U.S. National Institutes of Health (2005b) Imatinib mesylate with or without radiation therapy in treating young patients with newly diagnosed or recurrent glioma.
  40. Uhrbom L, Hesselager G, Nister M, Westermark B (1998) Induction of brain tumors in mice using a recombinant platelet-derived growth factor B-chain retrovirus. Cancer Res 58(23):5275–5279PubMedGoogle Scholar
  41. Wilson AP (2000) Cytotoxicity and viability assays. In: Masters JRW (ed) Animal cell culture, 3rd edn. Oxford University, Oxford, pp. 175–219Google Scholar
  42. Yamada SM, Yamaguchi F, Morrison RS, Takahashi H, Teramoto A (1998) Inhibition of fibroblast growth factor receptor 1 expression in human glioblastoma cell contributes to the cell growth suppression. No To Shinkei 50(12):1101–1105PubMedGoogle Scholar
  43. Yu S, Pu P, Jiang D (1998) A study on the relationship among activity of PDGFBB autocrine loop, cell proliferation and apoptosis in human glioma cells. Zhonghua Bing Li Xue Za Zhi 27(5):352–355PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Dietmar Gross
    • 1
  • Günther Bernhardt
    • 1
  • Armin Buschauer
    • 1
    Email author
  1. 1.Institute of PharmacyUniversity of RegensburgRegensburgGermany

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