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. 1997 Jun;88(6):564–577. doi: 10.1111/j.1349-7006.1997.tb00420.x

Modulation of Motility and Proliferation of Glioma Cells by Hepatocyte Growth Factor

Shinji Yamamoto 1,2, Hiroaki Wakimoto 1,2, Masaru Aoyagi 2, Kimiyoshi Hirakawa 2, Hirofumi Hamada 1,✉
PMCID: PMC5921469  PMID: 9263534

Abstract

Invasive proliferation is a critical biological characteristic of gliomas. We evaluated the activities of hepatocyte growth factor (HGF) on proliferation and motility of glioma cells, comparing them with the effects of other growth factors (EGF, bFGF, PDGF‐BB, TGF‐βl). Seven primary culture lines all expressed c‐met and HGF mRNA, and secreted HGF. HGF stimulated 3H‐thymidine uptake of every glioma cell line (30 to 70% upregulation). Boyden chamber assay and scattering assay revealed that HGF promoted cell motility with chemokinetic and strong chemotactic activities. Concentric circle assay showed that HGF promoted two‐dimensional expansion (proliferation and motility) most strongly among the growth factors studied. Further, we analyzed 23 paraffin‐embedded sections of surgically resected gliomas (7 grade II, 8 grade III, and 8 grade IV) by immunohistochemistry. Expression of HGF and Met increased with malignant progression of gliomas, suggesting that gliomas stimulated their invasive proliferation by autocrine HGF production. Neurons and vasculature were HGF‐positive, and Met‐positive glioma cells gathered around them. The data indicate that neurons and vasculature, which are the main tracks of glioma invasion, augment chemotactic invasion and proliferation of gliomas by paracrine HGF secretion. Clearly HGF plays a critical role in invasive proliferation of glioma cells and it is therefore a candidate target of therapeutic intervention.

Keywords: Astrocytoma, HGF, Met, Invasion, Proliferation

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REFERENCES

  • 1. ) The Committee of Brain Tumor Registry of Japan . “Brain Tumor Registry of Japan ” ( 1993. ).
  • 2. ) Mahaley , M. S.Jr. , Mettlin , C. , Natarajan , N. , Laws , E. R. , Jr. and Peace , B. B.National survey of patterns of care for brain‐tumor patients . J. Neurosurg. , 71 , 826 – 836 . [DOI] [PubMed] [Google Scholar]
  • 3. ) Maxwell , H. P.The incidence of interhemispheric extension of glioblastoma multiforme through the corpus cal‐losum . J.Neurosurg. , 3 , 54 – 57 ( 1946. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Chicoine , M. R. and Silbergeld , D. L.The in vitro motility of human gliomas increases with increasing grade of malignancy . Cancer , 75 , 2904 – 2909 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Wakimoto , H. , Aoyagi , M. , Nakayama , T. , Nagashima , G. , Yamamoto , S‐ , Tamaki , M. and Hirakawa , K , Prognostic significance of Ki‐67 labeling indices obtained using MIB‐1 monoclonal antibody in patients with supraten‐torial astrocytomas . Cancer , 77 , 373 – 380 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Cerame , M. A. , Guthikonda , M. and Kohli , C. M.Ex‐traneural metastases in gliosarcoma: a case report and review of the literature . Neurosurgery , 17 , 413 – 418 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Lund‐Johansen , M. , Engebraaten , O. , Bjerkvig , R. and Laerum , O. D.Invasive glioma cells in tissue culture . Anticancer Res. , 10 , 1135 – 1152 ( 1990. ). [PubMed] [Google Scholar]
  • 8. ) De Clerck , Y. A. , Shimada , H. , Gonzalez‐Gomez , I. and Raffel , C.Tumoral invasion in the central nervous system . J. Neuro-Oncol , 18 , 111 – 121 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Rutka , J. T. , Hubbard , S. L. , Fukuyama , K. , Matsuzawa , K. , Dirks , P. B. and Becker , L. E.Effects of antisense glial fibrillaryacidicproteincomplementaryDNAonthe growth, invasion, and adhesion of human astrocytoma cells . Cancer Res. , 54 , 3267 – 3272 ( 1994. ). [PubMed] [Google Scholar]
  • 10. ) Merzak , A. , McCrea , S. , Koocheckpour , S. and Pilkington , G. J.Control of human glioma cell growth, migration and invasion in vitro by transforming growth factor β1 . Br. J. Cancer , 70 , 199 – 203 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. ) Pedersen , P.‐H. , Ness , G. O. , Engebraaten , O. , Bjerkvig , R. , Lillehaug , J. R. and Laerum , O. D.Heterogeneous response to the growth factors (EGF, PDGF(bb), TGF‐α, bFGF, IL‐2) on glioma spheroid growth, migration and invasion . Int. J. Cancer , 56 , 255 – 261 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Rubin , J. S. , Bottaro , D. P. and Aaronson , S. A , Hepato‐cyte growth factor/scatter factor and its receptor, the c‐met proto‐oncogene product . Biochim. Biophys. Acta , 1155 , 357 – 371 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Zarnegar , R. and Michalopoulos , G. K.The many faces of hepatocyte growth factor: from hepatopoiesis to he‐matopoiesis . J. Cell Biol , 129 , 1177 – 1180 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. ) Matsumoto , K. and Nakamura , T.Emerging multipotent aspects of hepatocyte growth factor . J. Biochem. , 119 , 591 – 600 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Watanabe , H. , Carmi , P. , Hogan , V. , Raz , T. , Silletti , S. , Nabi , I. R. and Raz , A.Purification of human tumor cell autocrine motility factor and molecular cloning of its receptor . J. Biol.Chem. , 266 , 13442 – 13448 ( 1991. ). [PubMed] [Google Scholar]
  • 16. ) Stracke , M. L. , Krutzsch , H. C. , Unsworth , E. J. , Arestad , A. , Cioce , V. , Schiffmann , E. and Liotta , L. A.Identification, purification, and partial sequence analysis of auto‐taxin, a novel motility‐stimulating protein . J. Biol. Chem. , 267 , 2524 – 2529 ( 1992. ). [PubMed] [Google Scholar]
  • 17. ) Murata , J. , Lee , H.Y. , Clair , T. , Krutzsch , H.C. , Arestad , A. A. , Sobel , M. E. , Liotta , L. A. and Stracke , M. L.cDNA cloning of the human tumor motility‐stimulating protein, autotaxin, reveals a homology with phos‐phodiesterases . J.Biol. Chem. , 269 , 30479 – 30484 ( 1994. ). [PubMed] [Google Scholar]
  • 18. ) Gaudino , G. , Follenzi , A. , Naldini , L. , Collesi , C. , Santoro , M. , Gallo , K. A. , Godowski , P. J. and Comoglio , P. M.Ronis a heterodimeric tyrosine kinase receptor activated by the HGF homologue MSP . EMBO J. , 13 , 3524 – 3532 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. ) Nakamura , T. , Nishizawa , T. , Hagiya , M. , Seki , T. , Shimonishi , M. , Sugimura , A. , Tshiro , K. and Shimizu , S.Molecular cloning and expression of human hepatocyte growth factor . Nature , 342 , 440 – 443 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Nakamura , Y. , Morishita , R. , Higaki , J. , Kida , L , Aoki , M. , Moriguchi , A. , Yamada , K. , Hayashi , S. , Yo , Y. , Matsumoto , K. , Nakamura , T. and Ogihara , T.Expression of local hepatocyte growth factor system in vascular tissues . Biochem. Biophys. Res. Commun. , 215 , 483 – 488 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Di Renzo , M. F. , Narsimhan , R. P. , Olivero , M. , Bretti , S. , Giordano , S. , Medico , E. , Gaglia , P. , Zara , P. and Comoglio , P. M.Expression of the Met/HGF receptor in normal and neoplastic human tissues . Oncogene , 6 , 1997 – 2003 ( 1991. ). [PubMed] [Google Scholar]
  • 22. ) Wullich , B. , Milller , H.‐W. , Fischer , U. , Zang , K.‐D. and Meese , E.Amplified met gene linked to double minutes in human glioblastoma . Eur.J. Cancer , 29A , 1991 – 1995 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Muleris , M. , Almeida , A. , Dutrillaux , A. M. , Pruchon , E. , Vega , F. , Delattre , J. Y. , Poisson , M. , Malfoy , B. and Dutrillaux , B.Oncogene amplification in human gliomas: a molecular cytogenetic analysis . Oncogene , 9 , 2717 – 2722 ( 1994. ). [PubMed] [Google Scholar]
  • 24. ) Moriyama , T. , Kataoka , H. , Tsubouchi , H. and Koono , M.Concomitant expression of hepatocyte growth factor (HGF), HGF activator and c‐met genes in human glioma cells in vitro . FEBS Lett , 372 , 78 – 82 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 25. ) Takahashi , M. , Ota , S. , Shimada , T. , Hamada , E. , Kawabe , T. , Okudaira , T. , Matsumura , M. , Kaneko , N. , Terano , A. , Nakamura , T. and Omata , M.Hepatocyte growth factor is the most potent endogenous stimulant of rabbit gastric epithelial cell proliferation and migration in primary culture . J. Clin. Invest. , 95 , 1994 – 2003 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. ) Horie , S. , Higashihara , E. , Nutahara , K. , Mikami , Y. , Okubo , A. , Kano , M. and Kawabe , K.Mediation of renal cyst formation by hepatocyte growth factor . Lancet , 344 , 789 – 791 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Shin , R. W. , Iwaki , T. , Kitamoto , T. and Tateishi , J.Methods in laboratory investigation: hydrated autoclave pretreatment enhances TAU immunoreactivity in formalin‐fixed normal and Alzheimer's disease brain tissue . Lab. Invest. , 64 , 693 – 702 ( 1991. ). [PubMed] [Google Scholar]
  • 28. ) Tashiro , K. , Hagiya , M. , Nishizawa , T. , Seki , T. , Shimonishi , M. , Shimizu , S. and Nakamura , T.Deduced primary structure of rat hepatocyte growth factor and expression of the mRNA in rat tissues . Proc. Natl. Acad. Sci. USA , 87 , 3200 – 3204 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. ) Di Renzo , M. F. , Bertolotto , A. , Olivero , M. , Putzolu , P. , Crepaldi , T. , Schiffer , D. , Pagni , C. A. and Comoglio , P. M.Selective expression of the Mer/HGF receptor in human central nervous system microglia . Oncogene , 8 , 219 – 222 ( 1993. ). [PubMed] [Google Scholar]
  • 30. ) Jung , W. , Castren , E. , Odenthal , M. , vande Woude , G. F. , Ishii , T. , Dienes , H.‐P. , Lindholm , D , and Schirmacher , P.Expression and functional interaction of hepatocyte growth factor‐scatter factor and its receptor c‐met in mammalian brain . J. Cell Biol , 126 , 485 – 494 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. ) Honda , S. , Kagoshima , M. , Wanaka , M. , Tohyama , K. , Matsumoto , K. and Nakamura , T.Localization and functional coupling of HGF and c‐Met/HGF receptor in rat brain: implication as neurotrophic factor . Brain Res. Mol, Brain Res. , 32 , 197 – 210 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Yamada , T. , Tsubouchi , H. , Daikuhara , Y. , Prat , M. , Comoglio , P. M. , McGeer , P. L. and McGeer , E. G.Immunohistochemistry with antibodies to hepatocyte growth factor and its receptor protein (c‐MET) in human brain tissue . Brain Res. , 637 , 308 – 312 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Hamanoue , M. , Takemoto , N. , Matsumoto , K. , Nakamura , T. , Nakajima , K. and Kohsaka , S.Neurotrophic effect of hepatocyte growth factor on central nervous system neurons in vitro . J. Neurosci. Res. , 43 , 554 – 564 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Moriyama , T. , Kataoka , H. , Seguchi , K. , Tsubouchi , H. and Koono , M.Effects of hepatocyte growth factor (HGF) on human glioma cells in vitro: HGF acts as a motility factor in glioma cells . Int. J. Cancer , 66 , 678 – 685 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 35. ) Rosen , E. M. , Laterra , J. , Joseph , A. , Jin , L. , Fuchs , A. , Way , D. , Witte , M. , Weinand , M. and Goldberg , I. D.Scatter factor expression and regulation in human glial tumors . Int. J. Cancer , 67 , 248 – 255 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 36. ) Landau , B. J. , Kwaan , H. C. , Verrusio , E. N. and Brem , S. S.Elevated levels of urokinase‐type plasminogen activator and plasminogen activator inhibitor type‐1 in malignant human brain tumors . Cancer Res. , 54 , 1105 – 1108 ( 1994. ). [PubMed] [Google Scholar]
  • 37. ) Yamamoto , M. , Sawaya , R , , Mohanam , S. , Bindal , A. K. , Bruner , J.M. , Oka , K. , Rao , V. H. , Tomonaga , M. , Nicolson , G. L. and Rao , J. S.Expression and localization of urokinase‐type plasminogen activator in human astrocy‐tomas in vivo . Cancer Res. , 54 , 3656 – 3661 ( 1994. ). [PubMed] [Google Scholar]
  • 38. ) Miyazawa , K. , Shimomura , T. , Kitamura , A. , Kondo , J. , Morimoto , Y. and Kitamura , N.Molecular cloning and sequence analysis of the cDNA for a human serine protease responsible for activation of hepatocyte growth factor . J. Biol Chem. , 268 , 10024 – 10028 ( 1993. ). [PubMed] [Google Scholar]
  • 39. ) Mizuno , K. , Inoue , H. , Hagiya , M. , Shimizu , S. , Nose , T. , Shimohigashi , Y. and Nakamura , T.Hairpin loop and second kringle domain are essential sites for heparin binding and biological activity of hepatocyte growth factor . J. Biol Chem. , 269 , 1131 – 1136 ( 1994. ). [PubMed] [Google Scholar]
  • 40. ) Dow , K. E. , Mirski , S. E. , Roder , J. C. and Riopelle , R. J.Neuronal proteoglycans: biosynthesis and functional interaction with neurons in vitro . J. Neurosci. , 8 , 3278 – 3289 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41. ) Steck , P. A. , Moser , R. P. , Bruner , J. M. , Liang , L. , Freidman , A. N. , Hwang , T. L. and Yung , W. K.Altered expression and distribution of heparan sulfate proteoglycans in human gliomas . Cancer Res. , 49 , 2096 – 2103 ( 1989. ). [PubMed] [Google Scholar]
  • 42. ) Rosenblurn , M. L.Factors influencing tumor cell traffic in the central nervous system . J. Neuro-Oncol , 17 , 175 – 177 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 43. ) Nagano , N. , Sasaki , H. , Aoyagi , M. and Hirakawa , K.Invasion of experimental rat brain tumor: early morphological changes following micromjection of C6 glioma cells . Acta Neuropathol. (Berl.) , 86 , 117 – 125 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 44. ) Hermanson , M. , Funa , K. , Hartman , M. , Claesson‐Welsh , L. , Heldin , C.‐H. , Westermark , B. and Nister , M.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 , 3213 – 3219 ( 1992. ). [PubMed] [Google Scholar]
  • 45. ) Berkman , R. A. , Merrill , M. J. , Reinhold , W. C. , Monacci , W. T. , Saxena , A. , Clark , W. C. , Robertson , J. T. , All , I. U. and Oldfield , E. H.Expression of the vascular permeabilityfactor/vascular endothelial growthfactor gene in central nervous system neoplasms . J. Clin. Invest. , 91 , 153 – 159 ( 1993. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46. ) Wakabayashi , Y. , Shono , T. , Isono , M. , Hori , S. , Matsushima , K. , Ono , M. and Kuwano , M.Dual pathways to tubular morphogenesis of vascular endothelial cells by human glioma cells: vascular endothelial growth factor/basic fibroblast growth factor and interleukin‐8 . Jpn. J. Cancer Res. , 86 , 1189 – 1197 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47. ) Bussolino , F. , Di Renzo , M. F. , Ziche , M. , Bocchietto , E. , Olivero , M. , Naldini , L.Gaudino , G. , Tamagnone , L. , Coffer , A. and Comoglio , P. M.Hepatocyte growth factor is a potent angiogenic factor which stimulates endothelial cell motilityand growth . J. Cell Biol , 119 , 629 – 641 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48. ) Rosen , E. M. , Grant , D. S. , Kleinman , H. K. , Goldberg , I. D. , Bhargava , M. M. , Nickoloff , B. J. , Kinsella , J. L. and Polverini , P.Scatter factor(hepatocytegrowth factor) is a potent angiogenesis factor in vivo . Symp. Soc. Exp. Biol , 47 , 227 – 234 ( 1993. ). [PubMed] [Google Scholar]
  • 49. ) Lang , F. F. , Miller , D. C. , Koslow , M. and Newcomb , E. W.Pathways leading to glioblastoma multiforme: a molecular analysis of genetic alterations in 65 astrocytic tumors . J.Neurosurg. , 81 , 427 – 436 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 50. ) Morrison , R. S. , Yamaguchi , F. , Bruner , J. M. , Tang , M. , McKeehan , W. and Berger , M. S.Fibroblast growth factor receptor gene expression and immunoreactivity are elevated in human glioblastoma multiforme . Cancer Res. , 54 , 2794 – 2799 ( 1994. ). [PubMed] [Google Scholar]
  • 51. ) Hermanson , M. , Funa , K. , Koopmann , J. , Maintz , D , Waha , A. , Westermark , B. , Heldin , C.‐H. , Wiestler , O. D. , Louis , D. N. , von Deimling , A. and Nistér , M.Association of loss of heterozygosity on chromosome 17p with high platelet‐derived growth factor α receptor expression in human malignant gliomas . Cancer Res. , 56 , 164 – 171 ( 1996. ). [PubMed] [Google Scholar]

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