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Hepatocyte growth factor stimulates the invasion of gallbladder carcinoma cell lines in vitro

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Abstract

Human gallbladder cancer is highly malignant and its prognosis is usually poor depending on the extent of surrounding tissue invasion. We examined in vitro the invasive activity of four gallbladder cancer cell lines (GB-d1, GB-h3, GB-d2 and FU-GBC-1) in the absence or presence of hepatocyte growth factor (HGF). In type 1 collagen gel culture, HGF stimulated cell proliferation and induced an invasive phenotype of arborizing structures in GB-d1, GB-h3 and GB-d2. In a Matrigel invasion assay, invasion was also induced in three of these cell lines by HGF but not in FU-GBC-1. Cellular motility was, however, stimulated by HGF in all of the four cell lines to various extents. Zymography for proteolytic enzymes demonstrated high levels of type IV collagenase and urokinase-type plasminogen activator (u-PA) activity in GB-d1, GB-h3 and GB-d2 even in the absence of HGF. In the presence of HGF, the 72 kDa type IV collagenase (MMP-2) activity of GB-h3 and u-PA activities of GB-d1, GB-h3 and GB-d2 were enhanced. In contrast, the MMPs and PAs activities of FU-GBC-1 were faint irrespective of the addition of HGF. A Western blot analysis demonstrated higher levels of 190 kDa c-MET product (HGF receptor) of GB-d1, GB-h3 and GB-d2 than that of FU-GBC-1. The invasion in the Matrigel assay stimulated by HGF was inhibited by protease inhibitors, aprotinin and FOY-305, as well as by anti-HGF antibody. These results thus suggest that, in addition to the importance of the proteolytic activity, the cellular motility induced via the HGF/HGF-receptor system is essential for the invasive progression of gallbladder carcinoma cells. © Rapid Science 1998

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References

  1. Henson DE, Albores-Saavedra J and Corle D, 1992, Carcinoma of the gallbladder: histologic type, stage of disease, grade, and survival rates. Cancer, 70, 1493–97.

    Google Scholar 

  2. Shimura H, Date K, Matsumoto K, Nakamura T and Tanaka M, 1995, Induction of invasive growth in a gallbladder cancer cell line by hepatocyte growth factor in vitro. Jpn J Cancer Res, 86, 662–9.

    Google Scholar 

  3. Shimura H, Date K, Miyazaki K and Tanaka M, 1993, Cell culture of biliary malignant tumors in collagen gel matrix. J Jpn Res Soc Gastroenterol Carcinog, 5, 375–9.

    Google Scholar 

  4. Nakamura T, Nishizawa T, Hagiya M, et al.1989, Molecular cloning and expression of human hepatocyte growth factor. Nature, 342, 440–3.

    Google Scholar 

  5. Seki T, Ihara I, Sugimura A, et al.1990, Isolation and expression of cDNA for different forms of hepatocyte growth factor from human leukocyte. Biochem Biophys Res Commun, 172, 321–7.

    Google Scholar 

  6. Rubin JS, Chan AM, Bottaro DP, et al.1991, A broad-spectrum human lung fibroblast-derived mitogen is a variant of hepatocyte growth factor. Proc Natl Acad Sci USA, 88, 415–19.

    Google Scholar 

  7. Bottaro DP, Rubin JS, Faletto DL, et al.1991, Identification of the hepatocyte growth factor receptor as the c-met protooncogene product. Science, 251, 802–4.

    Google Scholar 

  8. Naldini L, Vigna E, Narsimhan RP, et al.1991, Hepatocyte growth factor stimulates the tyrosine kinase activity of the receptor encoded by the proto-oncogene, c-met. Oncogene 6, 501–4.

    Google Scholar 

  9. Di Renzo MF, Narsimhan RP, Olivero M, et al.1991, Expression of the Met/HGF receptor in normal and neoplastic human tissues. Oncogene, 6, 1997–2003.

    Google Scholar 

  10. Weidner KM, Arakaki N, Hartmann G, et al.1991, Evidence for the identity of human scatter factor and human hepatocyte growth factor. Proc Natl Acad Sci USA, 88, 7001–5.

    Google Scholar 

  11. Stoker M, Gherardi E, Perryman M and Gray J, 1987, Scatter factor is a fibroblast-derived modulator of epithelial cell mobility. Nature, 327, 239–42.

    Google Scholar 

  12. Shima N, Nagao M, Ogaki F, Tsuda E, Murakami A and Higashio K, 1991, Tumor cytotoxic factor/hepatocyte growth factor from humar fibroblasts: cloning of its cDNA, purification and characterization of recombinant protein. Biochem Biophys Res Commun, 1801151–8.

    Google Scholar 

  13. Aoki Y, Shimura H, Li H, Date K and Tanaka M, 1995, Role of hepatocyte growth factor derived from human gallbladder fibroblasts in the invasion model of gallbladder cancer. J Jpn Res Soc Gastroenterol Carcinog, 7, 255–58.

    Google Scholar 

  14. Liotta LA, Rao CN and Barsky SH, 1983, Tumor invasion and the extracellular matrix. Lab Invest 49, 636–49.

    Google Scholar 

  15. Liotta LA and Stetler-Stevenson WG, 1991, Tumor invasion and metastasis: an imbalance of positive and negative regulation. Cancer Res, 51, 5054–59.

    Google Scholar 

  16. Woolley DE, 1984, Collagenolytic mechanisms in tumor cell invasion. Cancer Metastasis Rev, 3, 361–72.

    Google Scholar 

  17. Dano K, Andreasen PA, Grondahl-Hansen J, Kristensen P, Nielsen LS and Skriver L, 1985, Plasminogen activators, tissue degradation and cancer. Adv Cancer Res, 44, 139–266.

    Google Scholar 

  18. Johzaki H, Iwasaki H, Nishada T, Isayama T and Kikuchi M, 1989, A human gallbladder adenocarcinoma cell line. Cancer, 64, 2262–8.

    Google Scholar 

  19. Furukawa T, Kubota T, Watanaba M, et al.1992, Increased drug resistance of cultured human cancer cell lines in three-dimensional cellular growth assay using collagen gel matrix. J Surg Oncol, 49, 86–92.

    Google Scholar 

  20. Albini A, Iwamoto Y, Kleinman HK, et al.1987, A rapid in vitro assay for quantitating the invasive potential of tumor cell. Cancer Res, 47, 3239–45.

    Google Scholar 

  21. Mizoguchi H, Komiyama S, Matsui K, et al.1991, The response to epidermal growth factor of human maxillary tumor cells in terms of growth, invasion and expression of proteinase inhibitors. Int J Cancer, 49, 738–43.

    Google Scholar 

  22. Cajot JF, Sordat B, Kruithof EK and Bachmann F, 1986, Human primary colon carcinomas xenografted into nude mice. I. Characterization of plasminogen activators expressed by primary tumors and their xenografts. J Natl Cancer Inst, 77, 703–12.

    Google Scholar 

  23. Cajot JF, Sordat B and Bachmann F, 1986, Human primary colon carcinomas xenografted into nude mice. II. Modulation of tumor plasminogen activator activity by the host tissue environment. J Natl Cancer Inst, 77, 1099–107.

    Google Scholar 

  24. Sawaya RE, Yamamoto M, Gokaslan ZL, et al.1996, Expression and localization of 72 kDa type IV collagenase (MMP-2) in human malignant gliomas in vivo. Clin Exp Metastasis, 14, 35–42.

    Google Scholar 

  25. Tryggvason K, Hoyhtya M and Salo T, 1987, Proteolytic degradation of extracellular matrix in tumor invasion. Biochim Biophys Acta, 907, 191–217.

    Google Scholar 

  26. McDonnell S, Wright JH, Gaire M and Matrisian LM, 1994, Expression and regulation of stromelysin and matrilysin by growth factors and oncogenes. Biochem Soc Trans, 22, 58–63.

    Google Scholar 

  27. Stetler-Stevenson WG, Liotta LA and Kleiner DE Jr, 1993, Extracellular matrix 6: role of matrix metalloproteinases in tumor invasion and metastasis. FASEB J, 7, 1434–41.

    Google Scholar 

  28. Sato H, Kida Y, Mai M, et al.1992, Expression of genes encoding type IV collagen-degrading metalloproteinases and tissue inhibitors of metalloproteinases in various human tumor cells. Oncogene, 7, 77–83.

    Google Scholar 

  29. Rao JS, Steck PA, Tofilon P, et al.1993, Role of plasminogen activator and of 92-kDa type IV collagenase in glioblastoma invasion using an in vitroMatrigel model. J Neuro-Oncol, 18, 129–38.

    Google Scholar 

  30. Nakayama Y, Kohno K, Nomura Y, et al.1993, Enhanced invasive activity and decreased expression of tissue inhibitors of metalloproteinases by hepatocyte growth factor in human renal cancer cells. Cancer J, 6, 213–19.

    Google Scholar 

  31. Miyashita C, Wenzel E and Heiden M, 1988, Plasminogen: A brief introduction into its biochemistry and function. Haemostasis, 18, 7–13.

    Google Scholar 

  32. de Vries TJ, Quax PH, Denijn M, et al.1994, Plasminogen activators, their inhibitors, and urokinase receptor emerge in late stages of melanocytic tumor progression. Am J Pathol, 144, 70–81.

    Google Scholar 

  33. Duffy MJ, Reilly D, O'Sullivan C, O'Higgins N, Fennelly JJ and Andreasen P, 1990, Urokinase-plasminogen activator, a new and independent prognostic marker in breast cancer. Cancer Res, 50, 6827–9.

    Google Scholar 

  34. Sier CF, Verspaget HW, Griffioen G, Ganesh S, Vloedgravan HJ and Lamers CB, 1993, Plasminogen activators in normal tissue and carcinomas of the human oesophagus and stomach. Gut, 34, 80–5.

    Google Scholar 

  35. de Bruin PA, Griffioen G, Verspaget HW, et al.1988, Plasminogen activators profiles in neoplastic tissues of the human colon. Cancer Res, 48, 4520–4.

    Google Scholar 

  36. Sier CF, Verspaget HW, Griffioen G, et al.1991, Imbalance of plasminogen activators and their inhibitors in human colorectal neoplasia. Implications of urokinase in colorectal carcinogenesis. Gastroenterology, 101, 1522–8.

    Google Scholar 

  37. He C, Wilhelm SM, Pentland AP, et al.1989, Tissue cooperation in a proteolytic cascade activating human interstitial collagenase. Proc Natl Acad Sci USA, 86, 2632–6.

    Google Scholar 

  38. Mignatti P and Rifkin DB, 1993, Biology and biochemistry of proteinases in tumor invasion. Physiol Rev, 73, 161–95.

    Google Scholar 

  39. Naldini L, Tamagnone L, Vigna E, et al.1992, Extracellular proteolytic cleavage by urokinase is required for activation of hepatocyte growth factor/scatter factor. EMBO J, 11, 4825–33.

    Google Scholar 

  40. Naldini L, Vigna E, Bardelli A, et al.1995, Biological activation of pro-HGF (hepatocyte growth factor) by urokinase is controlled by a stoichiometric reaction. J Biol Chem, 270, 603–11.

    Google Scholar 

  41. Rosen EM and Goldberg ID, 1995, Regulation of scatter factor (hepatocyte growth factor) production by tumor-stroma interaction. EXS, 74, 17–31.

    Google Scholar 

  42. Jiang WG, Hallett MB and Puntis MC, 1993, Hepatocyte growth factor/scatter factor, liver regeneration and cancer metastasis. Br J Surg, 80, 1368–73.

    Google Scholar 

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Authors and Affiliations

  1. Department of Surgery 1, Kyushu University Faculty of Medicine, Fukuoka, Japan

    Hong Li, Hideo Shimura, Yasuaki Aoki, Kazuhiko Date & Masao Tanaka

  2. Division of Molecular Oncology, Biomedical Center, Osaka University Medical School, Osaka, Japan

    Kunio Matsumoto & Toshikazu Nakamura

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  1. Hong Li
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  2. Hideo Shimura
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  3. Yasuaki Aoki
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Li, H., Shimura, H., Aoki, Y. et al. Hepatocyte growth factor stimulates the invasion of gallbladder carcinoma cell lines in vitro. Clin Exp Metastasis 16, 74–82 (1998). https://doi.org/10.1023/A:1006516119518

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