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Molecular mechanism of proliteration and differentistion of male germ cells analyzed by using the cell line that undergoes "meiosis"in vitro.

Research Project

Project/Area Number 14370511
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Urology
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

FUJITA Jun  KYOTO UNIVERSITY Graduate School of Medicine, Professor, 医学研究科, 教授 (50173430)

Co-Investigator(Kenkyū-buntansha) NISHIYAMA Hiroyuki  KYOTO UNIVERSITY Graduate School of Medicine, Instructor, 医学研究科, 助手 (20324642)
ITOH Katsuhiko  KYOTO UNIVERSITY Graduate School of Medicine, Lecturer, 医学研究科, 講師 (90281097)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥9,800,000 (Direct Cost: ¥9,800,000)
Fiscal Year 2003: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥6,300,000 (Direct Cost: ¥6,300,000)
Keywordsmeiosis / germ cell / antibody / testis / ovary / tumor / 培養細胞 / モノクローナル抗体 / 免疫組織染色
Research Abstract

1. Using the GC-2M cell line, a clonal line derived from the mouse spermatocytic cell line GC-2spd(ts) and thought to undergo meiosis in vitro, we obtained the following results.
By RT-PCR, no expression of genes known to be specific to meiosis was detected in GC-2M. Karyotype analysis demonstrated that most cells have 74-75 chromosomes, near tetraploid, with some cells containing around 37 chromosomes, and that no haploid cells were ~present. Flowcytomery showed that the major peak in GC-2M as well as NIH/3T3 cells corresponded to 4n DNA contend Clonal culture of GC-2M cells with 4n or 8n DNA content suggested that they could proliferate and reduce the DNA content to 2n. These results indicate that the reduction of the number of chromosomes occurs in GC-2M cells, but the process is different from regular meiosis.
We raised mouse monoclonal antibodies against GC-2M. cell lysates, and obtained 10 clones. They immunohistochemically stained the sections of mouse testis. Trials to identify the proteins recognized by the antibodies were not successful.
2. We have established the first human ovarian dysgerminoma cell line HDGO. The karyotype was 46, XX, and expression of Vasa, a germ cell specific, protein, was detected in the cells. HDGO cells formed tumors in SCID mice, and constitutive activation of c-Kit proto-oncogene was identified.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Ohnishi, N. 他: "High expression of transgenes mediated by hybrid retroviral vectors"Gene Ther.. 9. 303-306 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagao, T. 他: "MAGE-A4 interacts with the liver oncoprotein gankyrin"J.Biol.Chem. 278. 10668-10674 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Higuchi, T. 他: "Cyclic AMP enhances the expression of an extravillous trophoblast marker"Mol.Hum.Reprod.. 9. 359-366 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kanamori, T. 他: "PEP-19 overexpression in human uterine leiomyoma"Mol.Hum.Reprod.. 9. 709-717 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakurai, T. 他: "A cleaved form of MAGE-A4 binds to Miz-1 and induces apoptosis"J.Biol.Chem. 279. 15505-15514 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nishiyama, H. 他: "Clinical outcome of a large-scale multi-institutional retrospective study"Eur.Urol.. 45. 176-181 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 藤田潤: "みんな知りたい遺伝のはなし"京都新聞出版センター. 95 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Higashitsuji, H., Higashitsuji, H., Nagao, T., Nonoguchi, K., Fujii S., Itoh, K., Fuiita, J.: "A novel protein overexpressed in hepatoma accelerates export of NF-κB from the nucleus and inhibits p53-dependent apoptosis."Cancer Cell. 2. 333-346 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ohnishi, N., Itoh.K, Itoh, Y., Baum, C., Higashitsuji, H., Yarnaguchi, K., Tsuji, T., Okanoue, T., Fujita J.: "High expression of transgenes mediated by hybrid retroviral vectors in hepatocytes : comparison of promoters from murine retroviruses in vitro and in vivo."Gene Ther.. 9. 303-306 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Fuhuhara, K., Kariya, M., Kita, M., Shime, H., Kanamori, T., Kosa a, C., Orii, A., Fuiita, J., Fujii, S.: "Secreted frizzled related protein 1 is overexpressed in uterine leiomyomas, associated with a high estrogenic environment and unrelated to proliferative activity."J.Clin.Endocrinol.Metab.. 87. 1729-1736 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagao, T., Higashitsuji, H., Nonoguchi, K, Sakurai, T., Dawson, S, Mayer, R.J., Itoh, K., Fujita, J.: "MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity."J.Biol.Chem.. 278. 10668-10674 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Yamaguchi, K., Itoh, K, Ohnishi, N., Itoh, Y, Baum, C., Tsuji, T., Nagao, T., Higashitsuji, H., Okanoue, T., Fujita, J.: "Engineered long terminal repeats of retroviral vectors enhance transgene expression in hepatocytes in vitro and in vivo"Mol.Ther.. 8. 796-803 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Higuchi, T., Fujiwara, H., Egawa, H., Sato, Y., Yoshioka, S., Tatsurni, K., Itoh, K., Maeda, M., Fujita, J., Fujii, S.: "Cyclic AMP enhances the expression of an extravillous trophoblast marker, melanoma cell adhesion molecule, in choriocarcinoma cell JEG3 and human chorionic villous explant cultures."Mol.Human Reprod.. 9. 359-366 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kanamori, T., Takakura, K., Mandai, M., Kariya, M., Fukuhara, K., Kusakari, T., Momma, C., Shime, H., Yagi, H., Konishi, M., Suzuki, A., Matsumura, N., Nanbu, K., Fujita, J., Fujii, S.: "PEP-19 Overexpression in Human Uterine Leiornyoma."Mol.Human Reprod.. 9. 709-717 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakurai, T., Itoh, K., Higashitsuji, H., Nagao, T., Nonoguchi, K., Chiba, T., Fujita, J.: "A cleaved form of MAGE-A4 binds to Miz-1 and induces apoptosis in human cells."J.Biol.Chem.. 279. 15505-15514 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nishiyama H, Habuchi T, Watanabe J, Teramukai S, Tada H, Ono Y, Ohshima S, Fujimoto K, Hirao Y, Fukushima M, Ogawa O.: "Clinical outcome of a large-scale multi-institutional retrospective study for locally advanced bladder cancer in Japan: A survey including 1131 patients treated during 1990-2000."Eur.Vrol.. 45. 176-181 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakurai, T.: "A cleaved form of MAGE-A4 binds to Miz-1 and induces apoptosis in human cells"J.Biol.Chem.. (印刷中). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Wellmann, S.: "Oxygen-regulated expression of the RNA-binding proteins RBM3 and CIRP by a HIF-1-independent mechanism"J.Cell Sci.. (印刷中). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nishiyama, H.: "Clinical outcome of a large-scale multi-institutional retrospective study for locally advanced bladder cancer : a survey including 1131 patients treated during/1990-2000 in Japan"Eur.Urol.. 45・2. 176-181 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kanamori, T: "PEP-19overexpression in human uterine leiomyoma"Mol.Hum.Reprod.. 9. 709-717 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Higuchi, T.: "Cyclic AMP enhances the expression of an extravillous trophoblast marker melanoma cell adhesion molecule, in choriocarcinoma cell JEG3 and human chorionic vilous explant cultures"Mol.Hum.Reprod.. 9. 359-366 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hamid, A.A.: "Expression of cold-inducible RNA-binding protein in the normal endometrium, endometrial hyperplasia and endometiral carcinoma"Int.J.Gynecol.Pathol.. 22. 240-247 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nagao, T.: "MAGE-A4 interacts with the liver oncoprotein gankyrin and suppresses its tumorigenic activity"J. Biol. Chem.. (印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Higashitsuji, H.: "A novel protein over expressed in hepatoma accelerates export of NF-KB from the nucleus and inhibits p53-dependent apoptosis"Cancer Cell. 2. 333-346 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Dawson, S: "Gankyrin : an Ankyrin-repeat Oncoprotein interacts with CDK4 Kinase and the S6 ATPase of the 26 S proteasome"J. Biol. Chem.. 277. 10893-10902 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ohnishi, N.: "High expression of transgenes mediated by hybrid retroviral vectors in hepatocytesi comparison of promoters from munine retroviruses in vitro and in vivo"Gene Ther.. 9. 303-306 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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