1987 Fiscal Year Final Research Report Summary
High resolution chromosomal analysis by the synchronization culture monitored with the flow cytometry for human hematological malignancies
| Project/Area Number |
61570580
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hematology
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| Research Institution | HIROSHIMA UNIVERSITY |
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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)
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| 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)
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| Project Period (FY) |
1986 – 1987
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| Keywords | Chromosomal abnormalities / High resolution chromosomal analysis / Methotrexate / Ethidium bromide / Flow cytometory / Acute leukemia / 骨髄異形成症候群 |
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| 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
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