| Project/Area Number |
06557078
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | Niigata University |
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Principal Investigator |
USUI Hiroshi Brain Res.Inst., Niigata Univ., Research Associate, 脳研究所, 助手 (20192510)
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| Co-Investigator(Kenkyū-buntansha) |
WASHIYAMA Kazuo Brain Res.Inst., Niigata Univ., Associate Professor, 脳研究所, 助教授 (00183715)
KUMANISHI Toshiro Brain Res.Inst., Niigata Univ., Professor, 脳研究所, 教授 (40018601)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥17,700,000 (Direct Cost: ¥17,700,000)
Fiscal Year 1996: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1995: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1994: ¥9,000,000 (Direct Cost: ¥9,000,000)
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| Keywords | brain tumor / tumor marker / gene expression / cDNA cloning / brain development / onco-fetal antigen / glioma / genemutation / onco‐fetal antigen / differential screening |
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| Research Abstract |
In this research project, we first established improved molecular biological procedures for efficient isolation of cDNA clones of differentially expressed mRNAs from small materials : (1) modified differential screening procedure, (2) directional tag PCR subtraction, an efficient subtractive cDNA cloning procedure, and (3) cDNA library DNA-Southern blot procedure which can replace Northern blot analysis.
Using these procedures, we successfully isolated 22 distinct rat cDNA clones whose corresponding mRNAs were preferentially expressed in the fetal brain during the brain development. We then isolated their human homologues and analyzed the mRNA expressions in various human tissues. We found that the expression pattern of the human neuronatin was notable. The neuronatin mRNA was selectively expressed in fetal human brain among normal human tissues and in human pituitary adenomas among various human brain tumors, suggesting the utilization as a new marker for the pituitary adenomas. We also analyzed the mRNA patterns of human glioma cell lines exhibiting different biological characters using the procedures described above. We identified 9 cDNA clones whose corresponding mRNAs were expressed more abundantly in the cells which showed hetero-transplantability and higher growth rate than the control cells. These cDNA clones included novel clones and those involved in the protein synthesis, signal transduction, and nucleotide metabolism. These findings showed that our improved procedures are useful to analyze molecular basis of tumor cells and to identify new tumor markers.
In addition, we analyzed gene mutations in human brain tumors to understand the molecular backgrounds of different mRNA expressions, and found frequent gene mutations of tumor suppressor genes, such as p53 and p16, in human brain tumors.
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