| Project/Area Number |
11670163
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Human pathology
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| Research Institution | Hirosaki University |
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Principal Investigator |
KUROTAKI Hidekachi HIROSAKI UNIVERSITY, SCHOOL OF MEDICINE, DEPARTMENT OF PATHOLOGY, ASSOCIATE PROFESSOR, 医学部, 助教授 (40215108)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGISHI Shin-ichiro HIROSAKI UNIVERSITY, SCHOOL OF MEDICINE, DEPARTMENT OF PATHOLOGY, INSTRUCTOR, 医学部, 助手 (80301026)
YAGIHASHI Soroku HIROSAKI UNIVERSITY, SCHOOL OF MEDICINE, DEPARTMENT OF PATHOLOGY, PROFESSOR, 医学部, 教授 (40111231)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | MICRODISSECTION / MYELODYSPLASTIC SYNDROME / CLONALITY / HUMARA / PGK / X-CHROMOSOME / PCR |
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| Research Abstract |
To explore the clonality of hematopoietic cells in myelodysplastic syndromes (MDS), we conducted clonal analysis by PCR-based phosphoglycerokinase gene (PGK) and human androgen receptor gene (HUMARA) assay in bone marrow specimens from 19 female patients with MDS (RA5, RARS 1, RAEB 13) and 5 female patients with de novo AML.After immunohistochemical staining for serial paraffin sections, laser capture microdissection was carried out on the immunostained sections and DNA was extracted from the sample from immunopositive cells. We employed anti-CD34 antibody for bone marrow progenitor cells, anti-glycophorin A antibody for erythropoietic cells, anti-myeloperoxidase antibody for granulopoietic cells and anti-glycoprotein IIb/IIIa antibody for megakaryopoietic cells. DNA samples were subjected to enzymatic digestion with HpaII or HhaI.PCR products were considered to be monoclonal when only one PCR product from the sample with enzyme digestion was observed on the gel electrophoresis. PGK analysis revealed the monoclonality of granulopoietic cells in 3 cases of RAEB and 2 cases of AML, and of progenitor cells in 4 cases of RAEB and 2 cases of AML.HUMARA disclosed the monoclonal proliferation of granulopoietic cells in 4 cases of RAEB and 3 cases of AML, and of progenitor cells in 5 cases of RAEB and 3 cases of AML.No monoclonality was observed in cases of RA and RARS.We could not show the monoclonality of erythropoietic cells and megakaryopoietic cells in all examined cases. Monoclonal proliferation of progenitor cells in the early stage of MDS is likely to associate with the development of leukemia in MDS, thus providing a predictive value of leukemic transformation.
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