Development of The Quantitative Method of MYC Oncoprotein for Diagnosis of Childhood Tumors

开发用于诊断儿童肿瘤的 MYC 癌蛋白定量方法

基本信息

批准号：
02670465
负责人：
SHIMATAKE Hiroyuki
金额：
$ 1.47万
依托单位：
Toho University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670465/
关键词：
Childhood Tumor N-myc Protein Anti N-myc Protein Antibody Anti Pan-myc Antibody Neuroblastoma Retinoblastoma ELISA Method Fusion Protein Expression System in E. Coli 網膜芽腫抗ーpanーmyc抗体

项目摘要

The purpose of this research project is to develop the method to quantitate the N-myc protein with anti N-myc antibody ; the protein being encoded by the human N-myc oncogene, a member of the myc gene family, with an attempt to improve the diagnostic skills of neuroblastoma. Five essential tools for the above purpose have been developed in the year of 1990 : 1) N-myc specific antigen 2) pan-myc specific antigen 3) N-pan-myc specific antigen 4) anti N-myc antibody 5) anti pan-myc antibody. Immunological detection of the N-myc protein in neuroblastoma tissues consists of two groups of methods ; one with immunohistochemical method that can detect the N-myc protein within each cell of the tumor, and another with western blotting or ELISA that quantitate the total amount of the protein of the tissue but not of the cell. During the execution of the project, the "heterogeneous" expression of the N-myc protein has been noted ; this has led to a concern of the possible erroneous interpretation of the, ELISA for the quantitation of the N-myc protein.The year of 1991 has, therefore, . been devoted to study the expression of the N-myc protein in the individual cell of neuroblastoma. The effort has revealed the followings : 1) Although the amount of the N-myc protein varies depending on the phases of the cell cycle, individual variation surpasses rather than the phase of the cell cycle. 2) The "heterogeneous" pattern of the expression of the N-myc protein in the neuroblastoma tissue is not affected even though the total amount of the protein varies as a whole by the exogenous stimulation. The above observations guarantee the current attempt to develop the quantitation method with ELISA and western blotting of the N-myc protein of the tumor tissue.

本研究项目的目的是开发用抗N-myc抗体定量N-myc蛋白的方法；该蛋白质由人类 N-myc 癌基因（myc 基因家族的成员）编码，旨在提高神经母细胞瘤的诊断技能。 1990 年开发了用于上述目的的五种基本工具： 1) N-myc 特异性抗原 2) pan-myc 特异性抗原 3) N-pan-myc 特异性抗原 4) 抗 N-myc 抗体 5) 抗 pan -myc抗体。神经母细胞瘤组织中 N-myc 蛋白的免疫学检测由两组方法组成；一种是采用免疫组织化学方法，可以检测肿瘤每个细胞内的 N-myc 蛋白，另一种是采用蛋白质印迹或 ELISA 来定量组织而非细胞的蛋白总量。在项目执行过程中，人们注意到N-myc蛋白的“异质”表达；这引起了对 N-myc 蛋白定量 ELISA 可能错误解释的担忧。因此，1991 年。致力于研究N-myc蛋白在神经母细胞瘤个体细胞中的表达。这项工作揭示了以下内容： 1）尽管N-myc蛋白的量根据细胞周期的阶段而变化，但个体差异超越了细胞周期的阶段。 2)神经母细胞瘤组织中N-myc蛋白表达的“异质”模式不受影响，即使该蛋白的总量因外源刺激而整体变化。上述观察结果保证了目前开发肿瘤组织N-myc蛋白的ELISA和蛋白质印迹定量方法的尝试。