食管癌细胞核仁抗原单克隆抗体的制备及其特性的初步鉴定

【目的】制备抗食管癌细胞核仁抗原单克隆抗体 ,用于寻找肿瘤的标志物和癌的病理诊断。【方法】以食管癌组织核基质为抗原 ,采用足垫法免疫Balb/c小鼠 ,运用杂交瘤技术建立分泌特异性单抗的杂交瘤细胞株 ,制备食管癌细胞抗原片和食管癌冰冻切片 ,采用免疫组化检测并采用蛋白印迹法 (Westernblotting)分析 ,鉴定是否为特异性单克隆抗体。【结果】获得一细胞株 (1 7/E)能稳定分泌抗体的杂交瘤细胞株 ,1 7/E能与食管癌细胞和其他 10种癌细胞的核仁反应 ,不与正常食管上皮细胞核仁反应。Westernblotting分析表明 ,1 7/E与食管癌组织核基质 41ku处有一反应带 ,与正常食管组织核基质无反应。【结论】 1 7/E可能是对食管癌等多种肿瘤细胞核仁抗原的特异性单克隆抗体 ,为深入研究肿瘤细胞核仁抗原的生物学特性及肿瘤病理诊断提供了有效的制剂 ,并具开发的潜在价值。

ThePreparation of Monoclonal Antibody to Nucleolar Antigen of Esophageal Carcinoma and Preliminary Identification of Its Charateristics

Objective To prepare monoclonal antibody to nucleolar antigen of esophageal carcinoma for searching of the marker substances of neoplasms cell and pathological diagnosis of carcinomas. Methods Using nuclear matrix of esophageal carcinoma as antigen to immune Balb/c mice by inoculating into pads of the hind feethand, and hybridoma technique, a hybridoma cell line secreting specific monoclonal antibody was established. By Western blotting analysis and immunohistochemistry on smear of esophageal carcinoma cell lines and frozen sections of esophageal carcinoma, to identify specificity of the monoclonal antibody. Results A hybridoma cell line (1 7/E) stably secreting antibody was created. The 1 7/E antibody reacted positively with nucleoli of esophageal carcinoma and other ten kinds of carcinoma and negatively to nucleoli of normal esophageal epithelium. Western blotting anlysis showed that 1 7/E antibody reacted to nuclear matrix of esophageal carcinoma at the site of 41 ku as a reaction band and not to that of normal esophagus. Conclusions Antibody 1 7/E may be specific to nucleolar antigen of esophageal carcinoma and other carcinomas. The antibody is worthy for further study of biological charateristics of tumor nucleolar antigens and pathological diagnosis of malignancies, and may be of commercial value.