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Anti-metastatic gene therapy by human amniotic fluid-derived bikunin

Research Project

Project/Area Number 11671607
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Obstetrics and gynecology
Research InstitutionHamamatsu University School of Medicine

Principal Investigator

KOBAYASHI Hiroshi  Hamamatsu Univ. Sch. Of Med., Obstet. Gynecol., Assistant Prof., 医学部附属病院, 講師 (40178330)

Co-Investigator(Kenkyū-buntansha) SUZUKI Mitsuaki  Jichi Med. Univ., Obstet. Gynecol., Prof., 医学部, 教授 (50110870)
OOI Hidekazu  Hamamatsu Univ. Sch. Of Med., Obstet. Gynecol., Assistant, 医学部附属病院, 助手 (10283368)
NISHIGUCHI Tomizo  Hamamatsu Univ. Sch. Of Med., Obstet. Gynecol., Assistant Prof., 医学部附属病院, 講師 (30198452)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2001: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsMetastasis / Anti-metastatic agent / Cancer Invasion / Gene therapy / 転移
Research Abstract

Bikunin (bik), a Kunitz-type protease inhibitor, also known as urinary trypsin inhibitor, is proposed as a main participant in the inhibition of tumor cell invasion and metastasis, possibly through the direct inhibition of cell-associated plasmin activity and suppression of urokinase-type plasminogen activator (uPA) mRNA expression. In the present study, we transfected the human ovarian carcinoma cell line HRA, highly invasive cells, with an expression vector harboring a cDNA encoding for human bik. This study was designed to investigate the effect of bik overexpression and changes in tumor cell phenotype and invasiveness in the stably transfected clones. Bik gene transfection of HRA gave the following results : (1) transfection of HRA with the bik cDNA resulted in five variants stably expressing functional bik ; (2) bik^+ clones exhibited a significantly reduced uPA mRNA expression as compared with the parental cells ; (3) bikunin negatively regulates the ERK1/2 activity ; (4) secretion-blocking treatments of bik^+ clones abrogated bik-mediated suppression of ERK1/2 activation and uPA expression ; (5) the regulation of invasion seen in the HRA cells is mainly mediated by the uPA-plasmin-MMP-2 system ; (6) transfection of HRA with the bik gene significantly reduced invasion, but not proliferation, adhesion, or migrate relative to the parental cells ; and (7) animals with bik^+ clones induced reduced peritoneal dissemination and long term survival. We conclude that transfection of HRA cells with the bik cDNA constitutively suppresses ERK1/2 activation which results in inhibition of uPA expression and subsequently reduces dissemination of bik^+ clones.

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (62 results)

All Other

All Publications (62 results)

  • [Publications] Kobayashi H., et al.: "Urinary trypsin inhibitor levels in amniotic fluid of normal human pregnancy : decreased levels observed at parturition"Am. J. Obstet. Gynecol.. 180. 141-147 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Serum Hyaluronic Acid Levels During Pregnancy and Labor"Obstet. Gynecol.. 93. 480-484 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sun, GW., Kobayashi H.: "Localization and interaction of hyaluronic acid and inter-alpha-trypsin inhibitor in stimulated preovulatory rat ovaries"Acta Histochem. Cytochem.. 32. 65-71 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Immunolocalization of hyaluronic acid and inter-alpha-trypsin inhibitor in mice"Cell Tissue Res.. 296. 587-597 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Urinary trypsin inhibitor down-regulates hyaluronic acid fragment-induced prostanoid production in cultured human amnion"Mol. Hum. Reprod. 5;662-667,1999. 5. 662-667 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Identification of link protein during follicle development and cumulus cell cultures in rats"Endocrinol. 140;3835-3842,1999. 140. 3835-3842 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nishiguchi T, Kobayashi H.: "Upregulation of thrombomodulin antigen levels in U937 cells by combined stimulation with estradiol-17 beta and vitamin K2 (Menaquinone 4)"Semin. Thromb. Hemost.. 25. 509-517 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sun, GW., Kobayashi H.: "Expression of link protein during mouse follicle development"J. Histochem. Cytochem.47;1433-1442,1999. 47. 1433-1442 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Identification and characterization of a Kunitz-type proteinase inhibitor in ascites fluid from patients with ovarian carcinoma"Int. J. Cancer. 87(1),44-54,2000. 87. 44-54 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Human myometrial cells in culture express specific binding sites for urinary trypsin inhibitor"Mol. Hum. Reprod. 6;101-108,2000. 6. 101-108 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Clinical significance of combined use of macrophage colony-stimulating factor and sequamous cell carcinoma antigen as a selective"Gynecologic Oncology. 77;405-409,2000. 77. 405-409 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Identity of urinary trypsin inhibitor-binding protein with link protein"J. Biol. Chem.. 275. 21185-21191 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shu F, Kobayashi H., et al.: "Activated protein C suppresses tissue factor expression on U937 cells in the endothelial protein C receptor-dependent manner"FEBS Lett.. 477. 208-211 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sun, GW., Kobayashi H.: "Immunohistochemical study of annexin V expression in placentae of preeclampsia"Gynecologic & Obstetric Investigation. 49. 17-23 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Suppression of urokinase-type plasminogen activator expression from human ovarian cancer cells by urinary trypsin inhibitor"Biochim Biophys Acta 1481:310-316,2000. 1481. 310-316 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Invasive capacity of heterotopic endometrium"Gynecologic & Obstetric Investigation. 50. 26-32 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sakata M, Kobayashi H., et al.: "Ryudocan expression by luteinized granulosa cells is associated with the process of follicle atresia"Fertil. Steril.. 74. 1208-1214 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ishiguro K, Kobayashi H.: "Syndecan-4 deficiency impairs the fetal vessels in the placental labyrinth"Developmental Dynamics. 219. 539-544 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Suppression of urokinase expression and invasiveness by urinary trypsin inhibitor is mediated through inhibition of protein kinase C and"J Biol Chem.. 276. 2015-2022 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki M., Kobayashi H.: "Structure and function analysis of urinary trypsin inhibitor (UTI): Identification of binding domains and signaling property of UTI"Biochimica. Biophys. Acta. 1547;26-36,2001. 1547. 26-36 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Hirashima Y., Kobayashi H.: "Characterization of binding properties of urinary trypsin inhibitor to cell-associated binding sites on human chondrosarcoma cell line HCS-2/8"J. Biol. Chem.. 276. 13650-1365697 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Benson MD., Kobayashi H.: "Immunologic studies in presumed amniotic fluid embolism"Obstet. Gynecol.. 97. 510-514 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Excretion of bikunin and its fragments in the urine of patients with renal stones"J. Urol. 166:268-274,2001. 166. 268-274 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "A case of glassy cell carcinoma of the uterine cervix effectively responding to chemotherapy with paclitaxel and carboplatin"Anti-Cancer Drugs.. 12. 627-630 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., et al.: "Suppression of urokinase expression and tumor metastasia by bikunin overexpression"Human Cell.. 14. 233-236 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Suzuki K., Sugino D., Terao T.: "Urinary trypsin inhibitor levels in amniotic fluid of normal human pregnancy : ecreased levels observed at parturition"Am. J. Obstet. Gynecol. 180. 141-147 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Sun G.W., Tanaka Y., Kondo T, Terao T.: "Serum Hyaluronic Acid Levels During Pregnancy and Labor"Obstet. Gynecol. 93. 480-484 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sun G.W., Kobayashi H., Terano T.: "Localization and interaction of hyaluronic |acid and inter-alpha-trypsin inhibitor in stimulated preovulatory rat ovaries. Acta Histchem"Cytochem. 32. 65-71 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Sun G.W., Terao T.: "Immunolocalization of hyaluronic acid and inter-alpha-trypsin inhibitor in mice"Cell Tissue Res. 296. 587-597 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Sun G.W., Terao T.: "Urinary trypsin inhibitor down-regulates hyaluronic acid fragment-induced prostanoid production in cultured human amnion cells by inhibiting cycboxygenase-2 expression"Mol. Hum. Reprod. 5. 662-667 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Sun G.W., Hirashima Y., Terao T.: "Identification of link protein during follicle development and cumulus cell cultures in rats"Endocrinol. 140. 3835-3842 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Nishiguchi T., Kobayashi T., Sugimura M., Kobayashi H., Terao T.: "Upregulation of thrombomodulin antigen levels in U937 cells by combined stimulation with estradiol-17 beta and vitamin K2 (Menaquinone 4)"Semin. Thromb. Hemost. 25. 509-517 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sun G.W., Kobayashi H., Terao T.: "Expression of link protein during mouse follicle dewlopment"J. Histochem. Cytochem. 47. 1433-1442 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Hirashima Y., Sun G.W., Ohi H., Fujie M. and Terao T: "Identification and characterization of a Kunitz-type proteinase inhibitor in ascites fluid from patients with ovarian carcinoma"Int. J. Cancer. 87(1). 44-54 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Hirashima Y. and Terao T.: "Human myometrial cells in culture express specific binding sites for urinary trypsin inhibitor"Mol. Hum. Reprod. 6. 101-108 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Suzuki M., Tamaru N,. Kobayashi H., Ohwada M., Terao T., Sato I.: "Clinical significance of combined use of macrophage colony-stimulating factor and sequamous cell carcinoma antigen as a selective diagnostic marker for squamous cell carcinoma arising in mature cystic teratoma of the ovary"Gynecologic Oncology. 77. 405-409 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Hirashima Y., Sun G.W., Fujie M., Nishida T., Takigawa M., Terao T.: "Identity of urinary trypsin inhibitor-binding protein with link protein"J. Biol. Chem. 275. 21185-21191 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shu Fang, Kobayashi H., Fukudome K., Tsuneyoshi N., Kimoto M. and Terao T.: "Activated protein C suppresses tissue factor expression on U937 cells in the endothelial protein C receptor-dependent manner"FEBS Lett. 477. 208-211 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shu F., Sugimura M., Kanayama N., Kobayashi H., Kobayashi T., Terao T.: "Immunohistochemical study of annexin V expression in placentae of preeclampsia"Gynecologic & Obstetric Investigation. 49(1). 17-23 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Suzuki M., Sun G.W., Hirashima Y. and Terao T.: "Suppression of urokinase-type plasminogen activator expression from human ovarian cancer cells by urinary trypsin inhibitor"Biochim Biophys Acta. 1481. 310-316 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H.: "Invasive capacity of heterotopic endometrium"Gynecologic & Obstetric Investigation. 50(Suppl1). 26-32 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sakata M., Kobayashi H., Sun G.W., Mochizuki O., Takagi A. and Kojima T.: "Ryudocan expression by luteinized granulosa cells is associated with the process of follicle atresia"Fertil. Steril. 74. 1208-1214 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ishiguro K., Kadomatsu K., Kojima T., Muramatsu H., Nakamura E., Ito M., Nagasaka T., Kobayashi H., Kusugami K, Saito H. and Muramatsu T.: "Syndecan-4 deficiency impairs the fetal vessels in the placental labyrinth"Developmental Dynamics. 219. 539-544 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H., Suzuki M., Tanaka Y., Hirashima Y., and Terao T.: "Suppression of urokinase expression and invasiveness by urinary trypsin inhibitor is mediated through inhibition of protein kinase C and MEK/ERK/c-Jun-dependent signaling pathways"J. Biol. Chem. 276. 2015-2022 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mika Suzuki, Hiroshi Kobayashi, Yoshiko Tanaka, Yasuyuki Hirashima and Toshihiko Terao: "Structure and function analysis of urinary trypsin inhibitor (UTI) : Identification of binding domains and signaling property of UTI by analysis of truncated proteins. Biochimica"Biophys. Acta. 1547. 26-36 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Hirashima Y., Kobayashi H., Suzuki M., Tanaka Y., Kanayama N,. Fujie M., Nishida T., Takigawa M. and Terao T.: "Characterization of binding properties of urinary trypsin inhibitor to cell-associated binding sites on human chondrosarcoma cell line HCS-2/8"J. Biol. Chem. 276. 13650-13656 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Benson MD., Kobayashi H., Silver RK., Oi H., Greenberger PA. and Terao T.: "Immunologic studies in presumed amniotic fluid embolism"Obstet. Gynecol. 97. 510-514 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mika Suzuki, Hiroshi Kobayashi, Kageyama S., Shibata K., Fujie M. and Toshihiko Terao: "Excretion of bikunin and its fragments in the urine of patients with renal stones"J. Urol. 166. 268-274 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sekine M. and The Japanese Familial Ovarian Cancer Study Group (Kobayashi H.), et al.: "Localization of a novel susceptibility gene for familial ovarian cancer to chromosome 3p22-p25"Hum. Mol. Genetics. 10(13). 1421-1429 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sekine M. and The Japanese Familial Ovarian Cancer Study Group (Kobayashi H.): "Localization of a novel susceptibility gene for familial ovarian cancer to chromosome 3p22-p25"Hum. Mol. Genet. 10(13). 1421-1429 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Hirashima Y., Kobayashi H., Nishiguchi T., Miura K. and Kanayama N.: "A case of glassy cell carcinoma of the uterine cervix effectively responding to chemotherapy with paclitaxel and carboplatin"Anti-Cancer Drugs. 12. 627-630 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Oi H., Kobayashi H., Tokunaga N., Kobayashi T., Kanayama N. and Terao T.: "Sialyl Tn and Zinc Coproporphyrin-1 as sero-diagnostic markers for amniotic fluid emboism (AFE) : Retrospective analysis of 66 patients with established and potential AFE"5th World Congress of Perinatal Medicine, Barcelona, Spain, September 23-27. (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H.: "Suppression of urokinase expression and tumor metastasis by bikunin overexpression"Human Cell. 14(3)(Review article). 233-236 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sekine M. and The Japanese Familial Ovarian Cancer Study Group (Kobayashi H.): "Mutatioal analysis of BRCA1 and BRCA2 and clinicopathologic analysis of ovarian cancer in 82 ovarian cancer families: Two common founder mutations of BRCA1 in Japanese population"Clin Cancer Res. 7. 3144-3150 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Oi H., Kobayashi H., Tokunaga N., Kobayashi T., Kanayama N., Terao T.: "Sialyl Tn and zinc coproporphyrin-1 as sero-diagnostic markers for amniotic fluid embolism (AFE) : Retrospective analysis of 66 patients with established or potential AFE"Perinatology. 5^<th> World Congress of Perinatal Medicine. 901-904 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sakata M., Utsu M., Maeda M. and Kobayashi H.: "Ultrasonic placental shunt flow detection and postpartum color agar infusion into placental vessels in monochorionic twin pregnancies"The Woman and Child before, during and after pregnancy. Rome (Italy), May 22-25, 2001. Editor Ermelando V. Cosmi. Monduzzi Editore. (International Proceeding Division).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi H: "Identification and characterization of a Kunite-type protease inhibitor in …"Int.J.Cancer. 87. 44-54 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kobayashi H: "Human myometrial cells in culture express binding cites for UT1"Mel.Hum.Reprod.. 6. 101-108 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kobayashi H.: "Identity of UT1-binding protein with link protein"J.Bial.Chem.. 275. 21185-21191 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kobayashi H: "Suppression of UPA expression from human ovarian cancer cells by UT1"Biochim Biophyo Acta. 1481. 310-316 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kobayshi H: "Invasive capacity of heterotopic endometrium"Gynecol.Obstet.Invest. 50. 26-32 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Suzuki M: "Clinical significance of combined use of macrophhage colony-stimulating factor and …"Gynecol. Oncol. 77. 405-409 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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