Antiproliferative effect of trapidil, a platelet-derived growth factor antagonist, on a glioma cell line in vitro

Restricted access

✓ Platelet-derived growth factor (PDGF) is produced by glioma cells. However, there is heterogeneity among glioma cell lines in the production of PDGF. It has been demonstrated that U251MG cells produce a PDGF-like molecule while U105MG cells do not.

Trapidil, a specific antagonist of PDGF, competes for receptor binding with PDGF. Therefore, the inhibitory effect of trapidil on the proliferation of glioma cells was investigated in vitro using two glioma cell lines. At 100 µg/ml, trapidil significantly inhibited the proliferation of U251MG cells (which produce the PDGF-like molecule). At the same trapidil concentration, the proliferation of U105MG cells (which do not produce the PDGF-like molecule) was not inhibited. The inhibitory effect of trapidil was remarkable on Days 3 and 4 of culture. After 4 days of incubation, the proliferation of U251MG cells was 46% of the control preparation. Trapidil enhanced the antitumor effect of 3-((4-amino-2-methyl-5-pyrimidinyl)ethyl)-1-(2-chloroethyl)-1-nitro-sourea (ACNU) against U251MG cells. The enhancing effect was highest on Days 4 and 6 of culture. After 6 days of incubation in the presence of 100 µg/ml trapidil and 1 µg/ml ACNU, the proliferation of U251MG cells was 18% of the control preparation. These findings suggest that trapidil interrupts the autocrine loop at the PDGF and PDGF-receptor level and that combination therapy with trapidil and ACNU may be useful in the treatment of glioma.

Article Information

Address reprint requests to: Jun-ichi Kuratsu, M.D., Department of Neurosurgery, Kumamoto University Medical School, 1-1-1 Honjo, Kumamoto 860, Japan.

© AANS, except where prohibited by US copyright law.

Headings

Figures

  • View in gallery

    The antiproliferative effect of anti-platelet-derived growth factor on U251MG and U105MG cells. The growth of U251MG cells was suppressed while the growth of U105MG cells was not affected.

  • View in gallery

    The effects of trapidil in doses of 10 and 100 µg/ml on U251MG (left) and U105MG (right) cells. Trapidil (100 µg/ml) markedly inhibited the proliferation of U251MG cells while U105MG cells were only slightly affected.

  • View in gallery

    The antitumor effects of trapidil (100 µg/ml) on U251MG cells over time. The percent growth was calculated on Days 2, 4, and 6 using the MTT assay (see text).

  • View in gallery

    The antitumor effects of combined trapidil and ACNU on U251MG cells. The cells were incubated with ACNU alone (0.1 to 5.0 µg/ml, open circles), or ACNU plus trapidil (10 µg/ml, squares), or ACNU plus trapidil (100 µg/ml, solid circles).

  • View in gallery

    At 4 days after initiation of the cultures (arrow), the media containing only trapidil (100 µg/ml) or trapidil (100 µg/ml) plus ACNU (1 µg/ml) were removed and the cultures re-fed with medium containing only 10% fetal bovine serum (FBS). The number of cells treated with trapidil alone increased rapidly after the change of culture medium (open circles). In contrast, the number of cells treated with trapidil plus ACNU decreased after the change to medium containing 10% FBS (solid circles).

  • View in gallery

    The antitumor effect of combined trapidil plus ACNU on U251MG cells over time. The cells were incubated with ACNU alone (1 µg/ml, open circles) or with a combination of trapidil (100 µg/ml) and ACNU (1 µg/ml, solid circles).

References

  • 1.

    Bologa LDeugnier MAJoubert Ret al: Myelin basic protein stimulates the proliferation of astrocytes: possible explanation for multiple sclerosis plaque formation. Brain Res 346:1992031985Bologa L Deugnier MA Joubert R et al: Myelin basic protein stimulates the proliferation of astrocytes: possible explanation for multiple sclerosis plaque formation. Brain Res 346:199–203 1985

    • Search Google Scholar
    • Export Citation
  • 2.

    Coffey RJ JrLeof EBShipley GDet al: Suramin inhibition of growth factor receptor binding and mitogenicity in AKR-2B cells. J Cell Physiol 132:1431481987Coffey RJ Jr Leof EB Shipley GD et al: Suramin inhibition of growth factor receptor binding and mitogenicity in AKR-2B cells. J Cell Physiol 132:143–148 1987

    • Search Google Scholar
    • Export Citation
  • 3.

    Doolittle RFHunkapiller MWHood LEet al: Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor. Science 221:2752771983Doolittle RF Hunkapiller MW Hood LE et al: Simian sarcoma virus onc gene v-sis is derived from the gene (or genes) encoding a platelet-derived growth factor. Science 221:275–277 1983

    • Search Google Scholar
    • Export Citation
  • 4.

    Fontana AGrieder AArrenbrecht STet al: In vitro stimulation of glia cells by a lymphocyte-produced factor. J Neurol Sci 46:55621980Fontana A Grieder A Arrenbrecht ST et al: In vitro stimulation of glia cells by a lymphocyte-produced factor. J Neurol Sci 46:55–62 1980

    • Search Google Scholar
    • Export Citation
  • 5.

    Hosang M: Suramin binds to platelet-derived growth factor and inhibits its biological activity. J Cell Biochem 29:2652731985Hosang M: Suramin binds to platelet-derived growth factor and inhibits its biological activity. J Cell Biochem 29:265–273 1985

    • Search Google Scholar
    • Export Citation
  • 6.

    Kohler NLipton A: Platelets as a source of fibroblast growth-promoting activity. Exp Cell Res 87:2973011974Kohler N Lipton A: Platelets as a source of fibroblast growth-promoting activity. Exp Cell Res 87:297–301 1974

    • Search Google Scholar
    • Export Citation
  • 7.

    Kuratsu JEstes JEYokota Set al: Growth factors derived from a human malignant glioma cell line, U-251MG. J Neurooncol 7:2252351989Kuratsu J Estes JE Yokota S et al: Growth factors derived from a human malignant glioma cell line U-251MG. J Neurooncol 7:225–235 1989

    • Search Google Scholar
    • Export Citation
  • 8.

    Lim RTurriff DETroy SSet al: Glia maturation factor: effect of chemical differentiation of glioblasts in culture. Science 195:1951961977Lim R Turriff DE Troy SS et al: Glia maturation factor: effect of chemical differentiation of glioblasts in culture. Science 195:195–196 1977

    • Search Google Scholar
    • Export Citation
  • 9.

    Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth 65:55631983Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth 65:55–63 1983

    • Search Google Scholar
    • Export Citation
  • 10.

    Nister MLibermann TABetsholtz Cet al: Expression of messenger RNAs for platelet-derived growth factor and transforming growth factor-α and their receptors in human malignant glioma cell lines. Cancer Res 48:391039181988Nister M Libermann TA Betsholtz C et al: Expression of messenger RNAs for platelet-derived growth factor and transforming growth factor-α and their receptors in human malignant glioma cell lines. Cancer Res 48:3910–3918 1988

    • Search Google Scholar
    • Export Citation
  • 11.

    Ohnishi HKosuzume HYamaguchi Ket al: [Pharmacological properties of trapidil: comparison with other coronary vasodilators.] Nippon Yakurigaku Zasshi 76:4955031980 (Jpn)Ohnishi H Kosuzume H Yamaguchi K et al: [Pharmacological properties of trapidil: comparison with other coronary vasodilators.] Nippon Yakurigaku Zasshi 76:495–503 1980 (Jpn)

    • Search Google Scholar
    • Export Citation
  • 12.

    Ohnishi HYamaguchi KShimada Set al: A new approach to the treatment of atherosclerosis and trapidil as an antagonist to platelet-derived growth factor. Life Sci 28:164116461981Ohnishi H Yamaguchi K Shimada S et al: A new approach to the treatment of atherosclerosis and trapidil as an antagonist to platelet-derived growth factor. Life Sci 28:1641–1646 1981

    • Search Google Scholar
    • Export Citation
  • 13.

    Ross RGlomset JKariya Bet al: A platelet-dependent serum factor that stimulates the proliferation of arterial smooth muscle cells in vitro. Proc Natl Acad Sci USA 71:120712101974Ross R Glomset J Kariya B et al: A platelet-dependent serum factor that stimulates the proliferation of arterial smooth muscle cells in vitro. Proc Natl Acad Sci USA 71:1207–1210 1974

    • Search Google Scholar
    • Export Citation
  • 14.

    Rutherford RBRoss R: Platelet factors stimulate fibroblasts and smooth muscle cells. Quiescent in plasma serum to proliferate. J Cell Biol 69:1962031976Rutherford RB Ross R: Platelet factors stimulate fibroblasts and smooth muscle cells. Quiescent in plasma serum to proliferate. J Cell Biol 69:196–203 1976

    • Search Google Scholar
    • Export Citation
  • 15.

    Sensenbrenner MDelaunoy JPLabourdette Get al: Effects of brain extracts on the proliferation and the maturation of astroglial and oligodendroglial cells in culture. Biochem Soc Trans 10:4244261982Sensenbrenner M Delaunoy JP Labourdette G et al: Effects of brain extracts on the proliferation and the maturation of astroglial and oligodendroglial cells in culture. Biochem Soc Trans 10:424–426 1982

    • Search Google Scholar
    • Export Citation
  • 16.

    Sheffield WDKim SU: Myelin basic protein causes proliferation of lymphocytes and astrocytes in vitro. Brain Res 132:5805841977Sheffield WD Kim SU: Myelin basic protein causes proliferation of lymphocytes and astrocytes in vitro. Brain Res 132:580–584 1977

    • Search Google Scholar
    • Export Citation
  • 17.

    Sjölund MThyberg J: Suramin inhibits binding and degradation of platelet-derived growth factor in arterial smooth muscle cells but does not interfere with autocrine stimulation of DNA synthesis. Cell Tissue Res 256:35431989Sjölund M Thyberg J: Suramin inhibits binding and degradation of platelet-derived growth factor in arterial smooth muscle cells but does not interfere with autocrine stimulation of DNA synthesis. Cell Tissue Res 256:35–43 1989

    • Search Google Scholar
    • Export Citation
  • 18.

    Stein CALaRocca RVThomas Ret al: Suramin: an anticancer drug with a unique mechanism of action. J Clin Oncol 7:4995081989Stein CA LaRocca RV Thomas R et al: Suramin: an anticancer drug with a unique mechanism of action. J Clin Oncol 7:499–508 1989

    • Search Google Scholar
    • Export Citation
  • 19.

    Takamiya YKohsaka SToya Set al: Possible association of platelet-derived growth factor (PDGF) with the appearance of reactive astrocytes following brain injury in situ. Brain Res 383:3053091986Takamiya Y Kohsaka S Toya S et al: Possible association of platelet-derived growth factor (PDGF) with the appearance of reactive astrocytes following brain injury in situ. Brain Res 383:305–309 1986

    • Search Google Scholar
    • Export Citation
  • 20.

    Tiell MLSussman IIGordon PBet al: Suppression of fibroblast proliferation in vitro and of myointimal hyper-plasia in vivo by the triazolopyrimidine, trapidil. Artery 12:33501983Tiell ML Sussman II Gordon PB et al: Suppression of fibroblast proliferation in vitro and of myointimal hyper-plasia in vivo by the triazolopyrimidine trapidil. Artery 12:33–50 1983

    • Search Google Scholar
    • Export Citation
  • 21.

    Waterfield MDScrace GTWhittle Net al: Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature 304:35391983Waterfield MD Scrace GT Whittle N et al: Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature 304:35–39 1983

    • Search Google Scholar
    • Export Citation
  • 22.

    Westermark BWasteson Å: A platelet factor stimulating human normal glial cells. Exp Cell Res 98:1701741976Westermark B Wasteson Å: A platelet factor stimulating human normal glial cells. Exp Cell Res 98:170–174 1976

    • Search Google Scholar
    • Export Citation

TrendMD

Metrics

Metrics

All Time Past Year Past 30 Days
Abstract Views 57 57 2
Full Text Views 146 113 1
PDF Downloads 65 48 0
EPUB Downloads 0 0 0

PubMed

Google Scholar