| Project/Area Number |
02404047
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Dermatology
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| Research Institution | Gunma University |
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Principal Investigator |
HIDEKAZU Ishikawa Gunma University School of Medicine, Department of Dermatology, Professor, 医学部, 教授 (70008233)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Hiroaki Institute of Endocrinology, Gunma University, Department of Protein Chemistry, A, 内分泌研究所, 助教授 (50008611)
HORIUCHI Ryuya Institute of Endocrinology, Gunma University, Department of Pharmacology, Associ, 内分泌研究所, 助教授 (90008342)
OHNISHI Kazunori Gunma University School of Medicine, Department of Dermatology, Assistant Profes, 医学部, 講師 (60176948)
ISHIKAWA Osamu Gunma University School of Medicine, Department of Dermatology, Assistant Profes, 医学部, 講師 (90168188)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Growth factor / Neurofibroma / Neurofibromatosis / Competitive binding assay / レックリングハウゼン病 / ペプチド同定 |
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| Research Abstract |
We previously found that a dialysable, low-molecular-weight factor isolated from sera of patients with neurofibromatosis enhanced the cell growth and [ ^3H] thymidine uptake of neunrofibroma-derived fibroblasts (NF cells). The factor (neurofibroma fibroblast-stimulating factor. NFSF) was purified by HPLC. The purified factor showing a DNA-synthesis stimulation effect on NF cells was specifically bounds to the cells. Then, we tentatively measured NFSF concentrations in sera of patients by a competitive binding assay technique. As a result, NFSF was clearly detected in the sera of 6 of 9 adult patients with neurofibromatosis. In contrast, the factor was not detected in the sera of 2 infantile patients with pigmented macules alone or any of 5 healthy adult controls. 1. The amount of NFSF isolated from the sera of the patients was, however, too small to determine the construction of amino acid sequence. Then we purified the NFSF from the neurofibromos. We observed NFSF-activity in gel filtration fraction of the tumor extract. 2. Further, the determination of growth-stimulating activity with NF cell ws disclosed to be difficult due to shortened lifespan of NF cells in vitro. : Colony-forming efficencies (CFEs) decreased in both NF cells and dermal fibroblasts as assay cycles progressed. However, the cumulative cell generations of NF cells were approximately half that of the dermal fibroblasts. Then we tried to find the suitable established cell lines that respond to partially purified NFSF similar to NF cells among 3T3, T98, NRK, Glioma 2 and other cells. NRK cell responded to the factor in dose dependent manner. Now, we are purifying the neurofibroma-derived NFSF, the growth activity of which is determined with NRK cell.
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