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1999 Fiscal Year Final Research Report Summary

Investigation of the molecular mechanisms for the development of osteosarcoma using the retinoblastoma gene chimeric mice.

Research Project

Project/Area Number 10470306
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Orthopaedic surgery
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

TOGUCHIDA Junya  Kyoto University, Institute for Frontier Medical Sciences, Associate Professor, 再生医科学研究所, 助教授 (40273502)

Co-Investigator(Kenkyū-buntansha) SASAKI Masao  Kyoto University, Radiation Bioloby Center, Professor, 放射線生物研究センター, 教授 (20013857)
NAKAMURA Takashi  Kyoto University, Faculty of Medicine, Departmet of Orthop. Surg., Professor, 医学研究科, 教授 (10201675)
Project Period (FY) 1998 – 1999
Keywordsosteosarcoma / retinoblastoma gene / chimeric mice / transformation
Research Abstract

To investigate the role of the retinobastoma (Rb) gene in the process of oncogenesis, we have performed the in vitro transformation experiments using the Rb(-/-) osteoblast as a starting material. First, we have created the Rb chimeric mice consisted of Rb(+/+) and Rb(-/-) cells by using Rb(-/-) ES cells. The Rb(-/-) osteoblasts were isolated from Calvary and long bones from newborn chimeric mice, and several clones were established. The genotypes of these clonal cells were analyzed by the allelespecific PCR and Rb(-/-) clones were further studied. The level of alkaline phosphatase (ALP) and osteocalcin, which are considered to be markers for differentiated-osteoblasts, were relatively low, whereas the expression the cbfal and BMP2 genes were almost equivalent with those of the Rb(+/+) osteoblastic cells.
By the differentiation-induction experiments, these cells showed increased ALP activity and produced mineralized matrix, suggesting that the Rb deficiency has no remarkable effects for the differentiaotn process of osteoblasts. We found that these cells showed marked reduction of growth ability when the generation number was over 90, which is thought to be the life span of normal rodent cells, indicating that the Rb deficiency has no remarkable effects for the immortality. We are currently creating the tetracycline-regulated Rb expression system in these cells to further investigate the phenotypic differences of Rb(+/+) and Rb(-/-) osteoblasts, such as the response for growth factors.
On the other hand, to obtain fully transformed cells, the dominant negative mutant of the human p53 gene was transfected into these cells. Transformants showed persistent growth ability even after the 90ィイD1thィエD1 generations, suggesting that the mutant p53 confer the immortality to these cells. We are currently analyzing the in vitro and in vivo phenotypes as fully neoplastic cells of these transfectants.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Kanoe,H., et al.: "Ampkification of the CDK4 gene in sarcomas : Tumo* specificity and relationship with the RB gene mutation"Anticancer Res.. 18. 2317-2322 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kanoe,H., et al.: "Characteristics of genomic breakpoints in TLS-CHOP trans locations in lipsarcomas suggest the inbvement of Translin and topoisomerase * in the process of thanslocation"Oncogene. 18. 721-729 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakamura,I., et al.: "Associafion of the human NPPS gene with ossificafion of the posterior longitudinal ligament ot the spino (OPLL)"Hum. Genet.. 104. 492-497 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamamoto,H., et al.: "High incidence of SV40・like sequprces detedion in tumour and peripheral blood cells of Japanese osteosarcome patients"Br. J. Cancer. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ito,H., et al.: "Hedgehog sigraling molecules in bone marrow calls of the intial stage of fracure repair"Biochem. Biophys. Res. commun.. 262. 443-451 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tachibana,A. et al.: "The FANCA gene in Japanese Fanconi anemia : reports of eight novel mutations and analysis of soquena jariability"Hum. Mutat.. 13. 237-244 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kanoe, Hiroshi.: "Amplification of the CDK4 gene in sarcomas : tumor specificity and relationship with the RB gene mutation."Anticancer Res. 18. 2317-2322 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kanoe, Hiroshi.: "Characteristics of genomic breakpoints in TLS-CHOP translocations in liposarcoma suggest the involvement of Translin and topoisomerase II in the process of translocation."Oncogene. 18. 721-729 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakamura, Isao.: "Association of the human NPPS gene with ossification of he posterior longitudinal ligament of the spine (OPLL)."Hum. Genet.. 104. 492-497 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamamoto, Hiroshi.: "High incidence of SV40-like sequences detection in tumour and peripheral blood cells of Japanese osteosarcoma patients."Br. J. Cancer. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ito, Hiromu.: "Hedgehog signaling molecules in bone marrow cells at the initial stage of fracture repair."Biochem. Biophys. Res. Commun.. 262. 443-451 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tachibana, Akira.: "The FANCA gene in Japanese Fanconi anemia : reports of eight novel mutations and analysis of sequence variability."Hum. Mut.. 13. 237-244 (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2001-10-23  

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