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骨組織由来腫瘍細胞誘引因子の単離・同定-ラット骨肉腫細胞株を用いて

Research Project

Project/Area Number 09770157
Research Category

Grant-in-Aid for Encouragement of Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Experimental pathology
Research InstitutionYokohama City University

Principal Investigator

長嶋 洋治  横浜市立大学, 医学部, 講師 (10217995)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywords転移 / 骨組織 / 骨肉腫 / 腎細胞癌 / SPARC / Osteonectin / BM-40 / 誘引因子 / Boyden chamber assay / IV型コラーゲン / 骨芽細胞 / BM40 / 遊走促進能 / 浸潤
Research Abstract

(背景)腎細胞癌、乳癌、前立腺癌などは骨転移の頻度が高く、末期に耐え難い苦痛を患者に与える。これらの骨高転移性を裏付ける分子機構を解明することは、患者のQuality of lifeの向上のみならず、骨転移の予防にもつながる。
(目的)本研究では、骨に由来する腫瘍細胞誘引因子の単離同定を行った。再現性を持って、多量の蛋白を抽出するために、ラット由来骨形成性骨肉腫細胞株YROS-1(Machida,Nagashima et al.,1995)の培養上清を用いた。
(方法)YROS-1の無血清培養上清を収集し、腎細胞癌細胞株を用いてのBoyden chamber assayを行い、腫瘍細胞誘引活性を検討した。次に、硫安沈殿、各種カラムを用いた分画化を行い、誘引活性が見られる分画に検討を加えた。Western blottingは型通り行った。
(結果)1. YROS-1の培養上清中には、腎細胞癌細胞株YCRに対する誘引活性が存在することがBoyden chamber assayで証明された。あらかじめ培養上清を細胞と混合することにより、同活性は中和された。
2. 硫安沈殿及びカラム操作にて、同活性を有する分画には、38kDaの分量を有する蛋白質が含まれていた。文献上、同蛋白質はSPARC/Osteonectin/BM-40と予想され、同蛋白に対する抗体を用いて行ったwestern blottingで反応性が確認された。また、中和抗体の存在下では、腫瘍細胞誘引活性は抑制された。
3. YROS-1培養上製の腫瘍紺胞誘引活性は、Boyden chamber膜がIV型コラーゲンにコートされている場合に上昇することが知られた。他の基質蛋白質では影響されなかった。
(結論)SPARCはIV型コラーゲンと結合することによって、少なくとも一部の腫瘍細胞に対する誘引活性を示す。この活性は腫瘍における骨高転移性を裏付ける分子機構である可能性がある。

Report

(2 results)
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Kato Y,Nagashima Y.et al.: "Stimulation of motility of human renal cell carcinoma by SPARC associated with type IV collagen." Invasion.Metastasis. (in press).

    • Related Report
      1998 Annual Research Report
  • [Publications] Taki A,Nagashima Y.et al.: "Chromophobe renal cell carcinoma.An immunohistochemical study of 21 Japanese cases." Modern Pathol. 12(3)(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nagashima Y et al.: "Pancreatoblastoma.A second report on cytogenetic findings." Cancer Genet Cytogenet. (in press).

    • Related Report
      1998 Annual Research Report
  • [Publications] Koshikawa N,Nagashima Y et al.: "Expression of trypsin by epithelial cells of various tissues,leukocytes and neurons in human and mouse." Am J Pathol. 153. 937-944 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shofuda K,Nagashima Y et al.: "Elevated expression of membrane-type 1 and 3 matrix metallo-proteinases in rat vascular smooth muscle cells." Lab Invest. 78. 915-923 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nagashima Y et al.: "Segmental mediolytic arteritis.A case report and review of the literature." Pathol Res Pract. 194. 643-647 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nagashima Y et, al.: "Expression of matrilysin in vascular endothelial cells adjacent to matrilysin-producing tumors." Int J Cancer. 72. 441-445 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kawano N, Nagashima Y et, al.: "Expression of gelatinase A, TIMP-2, matrilysin and trypsin (ogen) in lung neoplasms." Hum Pathol. 28. 613-622 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Hidai H, Nagashima Y, et al.: "Bilateral chromophobe renal carcinoma in tuberous sclerosis complex." Int J Urol. 4. 86-89 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Gorai I, Nagashima Y, et al.: "Uterine carcinosarcoma is derived from a single cell An in vitro study." Int J Cancer. 72. 821-827 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Koshikawa N, Nagashima Y, et al.: "Expression of trypsin vascular endothelial cells." FEBS Lett. 409. 442-448 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nagashima Y, et al.: "Adult ganglioneuroblastoma of the anterior mediastinum." Pathol Res Pract. 193. 727-732 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Takeuchi N, Nagashima Y, et al.: "Matrilysin gene expression in sporadic and familial colorectal. adenomas." Mol Carcinogenesis. 19. 225-229 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ishizuka S, Nagashima Y, et al.: "Regulation and immunohistochemical analysis of stress protein HO-1 in rat kidney with myoglobulinuric ARF." Biochem Biophys Res Comm. 240. 93-98 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Udagawa K, Nagashima Y, et al.: "Specific expression of pp5/TFPI-2 mRNA by syncytiotrophoblasts in human placenta as revealed by ISH." Placenta. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Nakatani Y, Nagashima Y, et al.: "Pulmonary adenocarcinoma of fetal lung type : A clinstudy indicating differrence." Am J Surg Pathol. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato Y, Nagashima Y, et al.: "Production of trypsin by human gastric cancer cells correlates with their malignant phenotypes." Eur J Cancer. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Ishizuka S, Nagashima Y, et al.: "Theophylline protects against glycerol-induced ARF Regulation of HO-1 and glutathione in rat kidney." Clin Exp Nephrol. 1. 204-211 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Numata M, Nagashima Y, et al.: "Inhibition of inducible NOS prevents LPS-induced acute lung injury in dogs." J Immunol. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Koshikawa N, Nagashima Y, et al.: "Expession of trypsin by endothelial cells of various tissues, leukocytes and neurons in mouse and human." Am J Pathol. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Momiyama N, Nagashima Y, et al.: "Inhibitory effect of matrilysin antisense oligonucleotides on human colon cancer invasion in vitro." Int J Cancer. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] 神座慎一郎, 長嶋洋治, 他: "浸潤性発育を示したBellini管癌の1例" 泌尿紀要. 43. 291-294 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 長嶋洋治, 他: "腎病変とvon Hippel-Lindau病" 病理と臨床. 15. 124-128 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 宮崎香、長嶋洋治他: "大腸癌の現在と未来-遺伝子診断の臨床形態学への応用-" メジカルビュー社(印刷中),

    • Related Report
      1997 Annual Research Report
  • [Publications] 長嶋洋治: "病理学実践マニュアル" 医学書院(印刷中),

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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