| Project/Area Number |
61480138
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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 |
Experimental pathology
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| Research Institution | Kobe University School of Medicine |
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Principal Investigator |
SUGIYAMA Taketoshi Kobe University School of Medicine. Professor, 医学部, 教授 (20030851)
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| Co-Investigator(Kenkyū-buntansha) |
KITAZAWA Souhei Kobe University School of Medicine. Assistant, 医学部, 助手 (90186239)
TAKAHASHI Rei Kobe University School of Medicine. Associate Professor, 医学部, 講師 (60144565)
MAEDA Sakan Kobe University School of Medicine. Associate Professor, 医学部, 助教授 (50030911)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | monoclonal antibody / gene product / pathology / ras / abl / B-11 / insulin / 分子異常 / 組織化学 / ホルモン / 甲状腺癌 |
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| Research Abstract |
Many kinds of monoclonal antibodies(MoAbs) are widely used for routine pathological diagnosis. However, MoAds against oncogenes are new tools and their specicicity and usefulness are still contro-versial. Although MoAbs against c-Ha-ras protein p21 had been reported to be specifically positive for cancer cells, we confirmed those MoAbs were positively reacted with both cancer and normal cells by the western blot method. Parallel relationship was also confirmed between the immuno-histochemical staining and the expression in messenger RNA detected by northern blot method. In another project, MoAbs against synthetic oligopeptides corresponding partially to v-abl protein sequence were generated in our laboratory and the abl-protein products were shown to localize on cell membrane by electron microscope. MoAbs were also generated by the immunization of human squamous cell or renal cell carci-nomas. B-11 MoAb detected 34kd protein and was reacted with squamous and transitional cell carconoma
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s but not with glandular cell carcinomas. When the grade of malignance of human bladder cancer increased, the positivity for B-11 decreased. Methodological improvement was made by using non-isotope labeled DNA probe for in situ hybridization. 5'-bromodeoxyuridine(brdurd) was incorporated into plasmid DNA by ino-culating E.coli and after hybridization with BrdUrd labeled DNA probe, the hybridized probe was detected by FITC-conjugated monoclonal antibody. Polymorphism in the 5'-flanking region of the human insulin gene in Japanese non-insulin-dependent diabetic subjects was analyzed and lower frequency of 5'-flanking in-sertion was observed than in non-Japanese reaces. We also analysed the mechanism of DNA degradation with formalin fixation in order to use the fixed samples for analysis of insulin or other genes. Use of buffered formalin at low temperature and EDTA were shown to block DNA degradation.
Further study is necessary to understand the usefulness and specificity of MoAbs against oncogenes and other genes products for molecular pathological studies of cellular process. Less
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