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High resolution chromosomal analysis by the synchronization culture monitored with the flow cytometry for human hematological malignancies

Research Project

Project/Area Number 61570580
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Hematology
Research InstitutionHIROSHIMA UNIVERSITY

Principal Investigator

OGUMA NOBUO  Research Institute for Nuclear Medicine & Biology, Hiroshima Universty. Associate Professor, 原爆放射能医学研究所, 助教授 (10034646)

Co-Investigator(Kenkyū-buntansha) KURAMOTO Atsushi  Research Institute for Nuclear Medicine & Biology, Hiroshima University. Profess, 原爆放射能医学研究所, 教授 (50034070)
IMAMURA NOBUTAKA  Research Institute for Nuclear Medicine & Biology, Hiroshima University. Lecture, 原爆放射能医学研究所, 講師 (60110821)
Project Period (FY) 1986 – 1987
Project Status Completed (Fiscal Year 1987)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1987: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsChromosomal abnormalities / High resolution chromosomal analysis / Methotrexate / Ethidium bromide / Flow cytometory / Acute leukemia / 骨髄異形成症候群 / 高精度染色体分析法 / メソトレキセート / サイミジン / セルソーター
Research Abstract

High resolution chromosomes were good for the assignment of breakpoints, precise identifiaction of the segments involved in the structural rearrangement and evaluation of breakpoint regions for possible subtle edeletions or duplications.
The method to obtain the high resolution chromosomes with the combination of methotrexate(MTX)-synclonization technique and ethidium bromide(EBr)- inhibition technique of chromosome condensation was established.
The incubation time with 10_<-5>M thymidine were monitored by flow cytometry. The appropriate time for thymidine to obtain the long chromosomes varied widely form case to case. However, it situated between 41/4 hours and 53/4 hours. Sometimes we combined the above two points materials whenever there was enough sample for analysis. The concentration of EBr was examined with three different one. Ten micrograms per ml was appropriated for analysis. Less than that didn't bring enough chromosomal condensation and more than that damaged the chromosomes … More . The mitotic index of the MTX-synchlonization culture decreased a little compared with the control method. We examined 33 cases with hematological malignancy by the high resolution chromosome analysis according to the above conditions. They consisted of acute granulocytic leukemia(FAB classification : MI=4 cases, M2=10 cases, M3=5 cases, M4=6 cases, M5=1 case, M6=1 case), myelodysplastic syndrome(FAB classification: RAEB=3 cases) and acute lymphocytic leukemia (FAB classification : L2=3 cases). The rate of chromosomal abnormalities increased by the culture methods compared with the direct bone marrow preparations. The translocation between Nos. 8 and 21 was detected frequently with almost same rate by the different kind of methods. But 5_q- and the translocation between Nos. 15 and 17 were detected significantly with hight rate by the introduction of culture technique compared to the direct method.
The procedure to obtain the long chromosome (prometaphase and early metaphase) and the analysis of them is time consuming. So far this method appears to be difficult to be used for the routine cytogenetic study. However, this method is necessary to detect the precise chromosome region such as in situ chromosomal hybridization, subtle deletions and duplications. Less

Report

(2 results)
  • 1987 Final Research Report Summary
  • 1986 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] 小熊信夫: Pharma Medica. 4. 33-45 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 小熊信夫: 長崎医会誌. 61. 521-528 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 鎌田七男: 昭和61年度原爆障害症調査研究班報告書. 137-140 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 蔵本淳: 昭和61年度原爆障害症調査研究班報告書. 131-132 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Yoshimitsu,K.: Amer.J.Med.Genet.28. 211-214 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] 小熊信夫: 日本臨床. 45. 467-468 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] OGUMA, N.: "Chromosome abnormalities in leukemia." Pharma Medica. 4. 33-45 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Oguma, N.: "A-bomb related RAEB cases with chromosome abnormalities in bone marrow cells." Nagasaki Med. J.61. 521-528 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kamada, N.: "Chromosome aberrations of peripheral lymphocytes and bone marrow cells in atomic bomb survivors. (V) with psecial reference to transforming gene." Annual Report on Atomic Bomb Late Effect and Casualties in 1986. 137-140 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Kuramoto, A.: "Hematodysplasia among A-bomb survivors." Annual Report on Atomic Bomb Late Effect and Casualties in 1986. 131-132 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Yoshimitsu, K.: "Decreased superoxide dismutase-2 activity in a patient with ring chromosome 6." Amer.J.Med.Genet.28. 211-214 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1987 Final Research Report Summary
  • [Publications] Nobutaka Imamura: American Journal of Hematology. 24. 115-116 (1987)

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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