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Production and characterization of newly developed anti-metastatic agents

Research Project

Project/Area Number 07671774
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Obstetrics and gynecology
Research InstitutionHAMAMATSU UNIV.SCH.OF MED.

Principal Investigator

KOBAYASHI Hiroshi  Hamamatsu Univ.Sch.Med.Dept.of Obstet.Gynecol.Assist.Professor, 医学部附属病院, 講師 (40178330)

Co-Investigator(Kenkyū-buntansha) NISHIGUCHI Tomizo  Hamamatsu Univ.Sch.Med.Dept.of Obstet.Gynecol.Assist.Professor, 医学部, 助手 (30198452)
TERAO Toshihiko  Hamamatsu Univ.Sch.Med.Dept.of Obstet.Gynecol.Professor, 医学部, 教授 (60022852)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Keywordsovarian cancer / invasion / metastasis
Research Abstract

Urinary trypsin inhibitor (UTI) efficiently inhibits tumor cell invasion and the formation of metastasis. The anti-metastatic effect is dependent on the COOH-terminal domain II of UTI (HI-8). In order to develop a molecule that binds with high affinity to the urokinase receptor (uPAR) on tumor cell surface, a bifunctional hybrid molecule (ATFHI) consisting of the uPAR-binding amino-terminal fragment (ATF) of uPA (amino acid sequence 1-134) at the NH_2-terminus of HI-8 was produced in Escherichia coli by genetic engineering. The purified hybrid protein inhibited trypsin and plasmin (inhibitory function was reduced two-to three-fold when compared with HI-8) and was also found to bind to human tumor cell via uPAR,which was confirmed by cell binding and competition experiments. Using a modified Boyden chamber and an artificial basement membrane, Matrigel, it was found that the hybrid protein is very effective at inhibiting invasion by uPAR-expressing human tumor cells. Sensitivities of tumor cells towards the anti-invasive effect of ATFHI correlated with the density of uPAR on human tumor cells. Furthermore, in the spontaneous metastasis model, the hybrid protein inhibited the formation of lung and /or lymphatic metastasis by human ovarian carcinoma and choriocarcinnoma cells. The hybrid protein was much more effective than ATF,HI-8, or UTI.We conclude that this approach extends the possibility of applying recombinant protein for therapeutic use in inhibition of human tumor cell metastasis.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Kobayashi H: "Effect of UTI on the invasion of reconstituted basement membarans by acarian cancer cells" Int.J.Cancer. 57. 378-384 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H: "UTI a kunitz-type potease inhibitor, ncsdulates tunnr necrosis factor-stimulated actration" Int.J.Oncol. 12. 95-105 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H: "Hyalimoni aid-sperifre regulation of cytobire by human ceteri" Am J Physiol. 273. 1151-1159 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H: "The role of cell-membrane associated UTI" Oncology Report. 4. 65-72 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H: "A bifunctional byhid potein of UPA-domain II" Recent Progress in Blood Congulation. 1. 249-252 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H: "|α| is concentrated in the pericellular environment of nonse gralers cells" Hum Pathol. 28. 428-433 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Fujie M,Shinohara H,Ohi H,Sugimura M,Terao T.: "Effect of urinary trypsin inhibitor on the invasion of reconstituted basement membranes by ovarian cancer cells." Int.J.Cancer.57. 378-384 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Fujishiro S,Terao T.: "Biochemical markers as a predictor for lymph node involvement in patients with cervical cancer of the uterus." Int.J.Oncol.4. 1053-1059 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Shinohara H,Takeushi K,Itho M,Fujie M,Saitoh M,Terao T.: "Inhibition of the soluble and the tumor cell receptorbound plasmin by urinary trypsin inhibitor and subsequent effects on tumor cell invasion and metastasis." Cancer Res.54. 844-849 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Moniwa N,Gotoh J,Sugimura M,Terao T.: "Role of activated protein C in facilitating basement membrane invasion by tumor cells." Cancer Res.54. 261-267 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Shinohara H,Ohi H,Sugimura M,Terao T,Hujie M.: "Urinary trypsin inhibitor (UTI) and fragments derived from UTI by limited proteolysis effciently inhibit tumor cell invasion.." Clin.Exp.Metastasis. 12. 117-128 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Gotoh J,Fujie M,Terao T.: "Chracterization of the cellular binding site for the urinary trypsin inhibitor." J.Biol.Chem. 269. 20642-20647 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H,Fujishiro S,Terao T.: "Impact of urokinase-type plasminogen activator and inhibitor type 1 on prognosis in cervical cancer of the uterus." Cancer Res.54. 6539-6545 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sugimura M,Kobayashi H,Terao T.: "Plasmin modulators, aprotinin and anti-catalytic plasmin antibody, efficiently inhibit destruction of bovine vascular endothelial cells by choriocarcinoma cells." Gynecol. Oncol.52. 337-346 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kobayashi H: "TYI,akunits-type potecase vlibitor,modulate TNF-α stinulated a ctretter and trancloutron of PKC in U93F alls" Int J Oncol. 12. 95-105 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi H: "Hyahronic acid-specitic regulation of cytokines by human uterie trabhroblests" Am J Physiol. 273. 1151-1159 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi H: "The role of cell-meubrace asseuated UTI in tamcll inasion and metastasis" oncol Report. 4. 65-72 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi H: "Histologic diafrosi of AFE on mAb TKH-2" Hum Pathol. 28. 428-433 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi H: "UTI efficiently inlibits UPA production" Em J Cell Biol. 71. 380-386 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi H: "CαL beud to tima cells in cleavel ato The HC and UTI." J Biol Chem. 271. 11362-11367 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kobayashi H: "Expression of urokinase and UTI" International Journal of Oncology. 8. 757-763 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kobayashi H: "Inter-α-trypsin inhibitor bound to tumor cells" Journal of Biological Chemistry. 271. 11362-11367 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kobayashi H: "UTI inhibits uPA production" Eurpean Journal of Cell Biology. 71. 380-386 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 小林浩: "Characterization of the cellular binding site for the urinary tryprin inhibity" J. Biol. Chem.,. 269. 20642-20647 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小林浩: "Inhibition of the soluble and the tama cell receptor-bound plssmin by urinany tuyprin nbibitor and sulsequent effect on tumur cell inrxsion" Cancer Res.,. 54. 844-849 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小林浩: "UTI and fraguarts diviued from UTI by limited poteolysis efficieutly inbibit tumy cell inrasies" Clin. Exp. Metastasis. 12. 117-128 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小林浩: "Effect of UTI on the inrasion of reconstituted basemant membranes by ovarian carca cells" Int. J. cancer. 57. 378-384 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小林浩: "Inhibitin of the wetastasis of Lews bey caruhova by awtibody against uPA in the expessweutal and eputaveon retastasis model" Thromb. Haemost.,. 71. 474-480 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] 小林浩: "Pole of activated proter C on facibitatby baseuovt membrane inrxsion by tumd cells." Cavcer Res.,. 54. 261-267 (1994)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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