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食管癌旁组织核基质蛋白单克隆抗体的制备及初步鉴定

Preparation of monoclonal antibodies to nuclear matrix proteins of tissues surrounding esophagus cancer and its preliminary identifi-cati on

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【作者】 张新梅林汉良刘乐和张昌卿钟叔平曾金云郑克立

【Author】 ZHANG Xin-mei; LIN Han-liang; LIU Le-he; ZHANG Chang-qing; ZHONG Shu-ping; ZENG Jin-yun; ZHENG Ke-li (Department of Microbiology, Sun Yat-sen University of Medical Scie nce, Guangzhou 510089, China; Department of Pathology, Sun Yat-se n University of Med

【机构】 中山医科大学微生物学教研室!广东广州510089中山医科大学病理学教研室!广东广州510089中山医科大学肿瘤医院中心实验室!广东广州510089汕头大学医学院生化教研室!广东汕头515000中山医科大学第一附属医院骨科!广东广州510089?

【摘要】 目的制备抗食管癌旁组织核基 质蛋白的特异性单抗。方法SDS-PAGE分析正常食管 组织、食管癌旁组织及食管癌组织核基质蛋白间的差异。用食管癌旁组织核基质蛋白做免疫 原免疫Balb/c小鼠,利用杂交瘤技术制备了5株能稳定分泌单克隆抗体的杂交瘤细胞株,并 对9-1-2/D、9-1-7/D 2株细胞作了初步鉴定。结果免疫组化分析发现9-1-2/D单抗与食管癌及正常食管组织均有反应,而9-1-7/D单抗 只与食管癌组织反应,与正常食管组织无反应。Western Blotting分析显示:9-1-2/D 单抗与电泳图谱上正常食管组织核基质和食管癌旁组织核基质52 kD的蛋白反应,而与食 管癌组织核基质55 kD的蛋白反应。9-1-7/D单抗只与电泳图谱上食管癌旁组织及癌组织 核基质46 kD的蛋白反应,与正常食管组织核基质无反应。结论本实验为食管癌的早期诊断作了初步有意义的探讨。

【Abstract】 ObjectiveTo prepare anti-nuclear ma trix proteins of tiss ues surrounding esophagus cancer monoclonal antibodies. Metho ds Differences among nuclear matri x proteins (NMPS) of normal e sophagus tissues, esophagus cancer tissues and tissues adjacent to can cer were identified by SDS-PAGE. Balb/c mouse were hyperimmunized wit h NMPS of tissues adjacent to esophagus cancer. Five mouse lymphocyte hybridoma cell lines were established and preliminary identification o f cell lines 9-1-2/D, 9-1-7/D were made.Results Immunohistochemical analysis showed monoclonal antibody 9-1-2/D reacte d both with esophagus cancer and normal esophageal tissues; monoclonal antibody 9-1-7/D reacted only with nucleoli of esophagus cancer cell s. Western Blotting analysis showed monoclonal antibody 9-1-2/D react ed with 52 kD NMPS of normal esophagus tissues and tissues adjacent to esophagus cancer and reacted with 55 kD NMP of esophagus cancer t issue. Monoclonal antibody 9-1-7/D react ed only with 46 kD protein of esophagus cancer tissues and tissues a djacent to cancer. Conclusion Monoclona l antibodies prepared by us might be a new method for early diagnosi s of esophagus cancer.

【基金】 1996年度广东省自然科学基金资助项目!(960777)
  • 【文献出处】 免疫学杂志 ,Immunological Journal , 编辑部邮箱 ,2000年06期
  • 【分类号】R735.1
  • 【被引频次】1
  • 【下载频次】31
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