| Project/Area Number |
09671826
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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 |
小児外科
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| Research Institution | Chiba University |
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Principal Investigator |
OHNUMA Naomi Chiba University, School of Medicine, Professor, 医学部, 教授 (50125910)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Hideo Chiba University, School of Medicine, Associate Professor, 医学部, 助教授 (60210712)
TNANABE Masahiro Chiba University, University Hospital, Professor, 医学部・付属病院, 教授 (10207160)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | neuroblastoma, / src / trk A / PCR / 神経芽種 / trk A / 神経特異的Src遺伝子 / N-myc / RT-PCR / 予後因子 / 神経特異的src遺伝子 |
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| Research Abstract |
To detect neuronal src mRNA expression in neuroblastoma specimens quickly after the removal, we tried to establish quantitative RT-PCR using β2-microglobulin as an internal marker. The cycles of PCR have been determined at the ranges, within which the PCR products of the target (src) and control (β2-microglobulin) genes were amplified in parallel according to the numbers of PCR cycles. The specficity of PCR products was confirmed by Southern blotting using src cDNA as a probe. The preliminary experiments using the 10 neuroblastoma cell lines, neuroblastoma cell line RT-BM-1 and its chemically differentiated cells, and 28 clinical samples from neuroblastomas indicated that the results from quantitative RT-PCR and S1 nuclease protection assay well corresponded. We also established quantitative RT-PCR for trk A expression. The expression of src and trk A genes in neuroblastoma tissues from 60 patients was examined. The tumors consisted of 24 at stage 1,4 at stage 2A, 7 at stage 2B, 5 at stage 3 and 20 at stage 4. Twelve patients died of the disease, all of which was at stage 4. N-myc gene amplification was observed in 9 tumors at stage 4, and all the patients with these tumors died. The results indicated that most of the tumors expressing c-srcN2 at the ratio more that 15% to total three c-src were at an early stage, and the prognosis was excellent. The 7 years survival rate of the patients with tumors expressing c-srcN2 at high ratio and low ratio was 93.8% and 35.3%, respectively (x2=24.519, P<0.0001). The combined analyses for c-srcN2 and trk A showed that the 7 years survival rate of the patients with tumors expressing both genes at high levels and low levels was 95.0% and 21.8%, respectively. The analyses for c-srcN2 and trk A expression by RT-PCR should provide accurate information about the biological phenotype of a neuroblastoma.
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