| Project/Area Number |
12670733
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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 |
Pediatrics
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| Research Institution | NIIGATA UNIVERSITY |
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Principal Investigator |
KAKIHARA Toshio NIIGATA university medical hospital, Associate professor, 医学部附属病院, 講師 (70262433)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Takehiro university medical hospital, Assistant, 医学部附属病院, 助手 (90311670)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2002: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | drug resistance / cell line / gene / leukemia / cloning |
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| Research Abstract |
The cDNA sequence of approximately 5,000 bases of a gene, which was increasedly expressed in drug-resistant leukemic cell line (KY-Ra), was determined with rapid amplication of cDNA ends method. A coding region comprised 357 bases was also determined
For evaluation of the function of the gene and relation to drug resistance, the expression of the gene in normal hematopoietic cells, leukemic cells and leukemic cell lines was quantitated using RT-PCR method. The increase of the gene expression was observed among normal hematopoietic cells, leukemic cells and leukemic cell lines in order, indicating the relation between the gene and cell proliferation and immortality. The gene was transfected into a cell line (HEK293) using expression vector (p3XFLAG). The stable transfectant showed resistance to anticancer drugs. The resistance was cross-resistance, suggesting the contribution to common pathway of apoptosis-inducing pathway. We are evaluating whether the required drug resistance described above occur in hematopoietic cells, and whether the gene was associated with other apoptosis-inducing pathway such as Fas-associated apoptosis. Furthermore, we are performing the purification of recombinant protein expressed in E.coli and the production of antibody against the protein for further evaluation.
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