| Project/Area Number |
08670177
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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 |
Pathological medical chemistry
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| Research Institution | Kobe University |
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Principal Investigator |
SANO Kimihiko Kobe University School of Medicine Department of Pediatrics, Assistant Professor, 医学部, 講師 (40205993)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Ecto-enzyme / Phosohodiesterase I / Nucleotide pyrophosphatase / Tumor metastasis / Motility / In situ hybridization / 神経芽細胞株 / 分子生物学 |
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| Research Abstract |
I have cloned two new members belonging to a phosphodiesterase I gene fimily from rat, and designated as PD-Ialpha and-beta. In current study I have cloned human PD-Ialpha and -beta and peroformed further analysis. I have found that human PD-Ialpha is an alternative splicing product of autotaxin, which is a newly found autocrine tumor cell motility-stimulating factor. I examined the expression of human PD-Ialpha/ATX gene in human neuroblastoma tumor tissues, motility stimulating activity of recombinant ATX on neuroblastoma cells, and investigated its transcriptional regulatory mechanism in a human neuroblastoma cell line. PD-Ialpha/ATX gene was expressed in the primary tumor tissues from the neuroblastoma patients at a various extent. This gene is also expressed in a SMS-KAN neuroblastoma cell line. We indentified both isoforms, PD-Ialpha and ATX,in these tumor tissues and SMS-KAN cells. The recombinant ATX stimulated the motility of SMS-KAN cells at low nanomolar concentration. We situated the promoter region, which is essential for its transcription in SMS-KAN cells, at-287 nt to -254 nt by the promoter activity assay. The gel shift assay revealed that there exists a nuclear protein in SMS-KAN cells that binds this region. These new insights about autocrine tumor cell motility-stimulating protein will help us to understand the metastatic mechanism of human neuroblastoma.
I have also cloned human PD-Ibeta. PD-Ibeta gene expression was mostly observed in the prostate and uterus. I have also shown that PD-Ialpha and beta are localized to human chromosome 8q24.1 and 6q22, respectively.
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