| Project/Area Number |
05454483
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
小児外科
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| Research Institution | TOHO UNIVERSITY |
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Principal Investigator |
SHIMATAKE Hiroyuki Toho University School of Medicine, Professor & Chairman, 医学部, 教授 (40010110)
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| Co-Investigator(Kenkyū-buntansha) |
MAKINO Shunichi Jichi Medical College, Associate Professor, 医学部, 助教授 (30157169)
SAJI Tsutomu Toho University School of Medicine, Associate Professor, 医学部, 助教授 (50120275)
INOUE Akira Toho University School of Medicine, Associate Professor, 医学部, 講師 (10151599)
HEMMI Hiromichi Toho University School of Medicine, Associate Professor, 医学部, 助教授 (90165514)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1994: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1993: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | myc oncogenes / N-myc oncogene / Anti N-Myc antibody / Anti pan-Myc antibody / Standwich-type ELISA / Neuroblastoma / Childhood tumor / 抗N-Myc抗体 / Myc系がん遺伝子 / N-Myc遺伝子 / 抗co-Myc抗体 / サンドウィッチ型ELISA / 小児がん |
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| Research Abstract |
Aberrant expression of human myc gene family members such as c-myc, N-myc, and L-myc occurs in many malignant tumors. Since N-myc gene amplification in neuroblastoma correlates its poor prognosis, amplification is believed to be important as a tumor marker. It is also important to quantitate N-Myc oncoprotein, because there is neuroblastoma of which N-myc gene is not amplified.
In this study, establishment of a quantification method for the gene products of myc gene family, especially N-Myc oncoprotein, and its clinical application were investigated. In first year (1994) supported by this grant, two polyclonal antibodies (anti N-Myc antibody and anti pan-Myc antibody) against N-Myc oncoprotein were prepared by the injection of antigens engineered by genetically. The specificity of these antibodies was studied by Western blot. Anti N-Myc antibody recognizes only N-Myc protein and anti pan-Myc antibody does N-Myc and c-Myc proteins. N-pan-Myc fusion protein was prepared by a similar procedure to that of the antigens and used as a standard protein.
In 1994, using the polyclonal antibodies and the standard protein, conditions for establishing an ELISA (enzyme-linked immunosorbent assay) system and preparation of monoclonal antibody were investigated. Three monoclonal antibodies were obtained by immunization with N-Myc fusion protein. However, these antibodies were not available for ELISA.Standard protein was detectable between 10 to 200 ng per ml. Then the contents of N-myc protein in human neuroblastoma cell lines were measured. Higher amount of the oncoprotein in neuroblastoma cells with amplification or without was detected than in a no N-myc expressing cell line.
Though it is required for an improvement for the ELISA system, it would be able to measure the N-Myc protein contents in clinical samples.
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