单克隆抗体N35对肺癌相关抗原的识别  

Characterization of lung cancer associated antigen by monoclonal antibody N 35

作　　者：王秦秦[1] 姜平[1] 李继梅[1] 唐睿珠[1] 陈新明[1] 

机构地区：[1]昆明医学院第一附属医院肿瘤研究所,云南昆明650032

出　　处：《中国癌症杂志》2000年第4期292-295,共4页China Oncology

基　　金：国家自然科学基金! ( 3 9960 0 78)

摘　　要：研究和确定肺癌相关抗原的有关特征及其意义。方法 :利用抗人肺癌单克隆抗体N 35作为免疫探针 ,经免疫沉淀和免疫印迹法测定其相关抗原在肺癌细胞系GLC 82、宫颈癌细胞系HeLa、肝癌细胞系HepG 2、乳腺癌细胞系PMC、正常人心脏及肺组织的存在及分布情况 ;用N 聚糖酶酶解方法确定肿瘤相关蛋白N35与糖蛋白分子的关系 ;用差速离心技术分离肺腺癌细胞系GLC 82亚细胞结构中的胞膜 ,胞核及线粒体成分 ,分别取样经免疫印迹法测定肿瘤相关抗原N35在亚细胞结构中的分布状况 ;用免疫荧光技术探测肺腺癌相关抗原N35在肿瘤细胞有丝分裂过程中的染色体结构定位 ;用 2种免疫亲和层析技术尝试获得大量纯化肿瘤相关蛋白N35的有效方法。结果 :肺腺癌相关抗原N35是一种糖蛋白分子 ,不存在于正常人心脏及肺组织蛋白组分中 ,而以不同Mr形式分布于GLC 82 ,Hela,HepG 2和PMC细胞蛋白中 ;在亚细胞结构中主要分布于胞核 ,线粒体次之 ,胞膜上分布最少 ,主要不以膜蛋白或跨膜蛋白的形式存在 ;在肿瘤细胞有丝分裂进入S至G2 期时明确地定位于中心粒 (centriole)结构上。结论 :肿瘤相关蛋白N35可能是一种只存在于肿瘤细胞并与其增殖活动密切相关的重要的肿瘤细胞生长调节蛋白 。Purpose:To identify and characterize lung cancer associated antigen detected by the monoclonal antibody N 35.Methods:Immunoprecipitation, immunoblotting,differential centrifugation and subcellular assay; N glycanase digestion;mitotic cell immunoflourescence and methods of affinity chromatography have been used by the monoclonal antibody N 35 as the immunoprobe to detect the distribution of lung cancer associated antigen N35 among various cancer cell lines and normal human tissue, the relationship between the lung cancer associated antigen and glycoprotein, the location of the subcellular structure and chromosomal domain of the lung cancer associated antigen, the most effective way for the purification of lung cancer associated antigen N35.Results:Lung cancer associated antigen N35 is a glycoprotein, distributed in the lung cancer cell line GLC 82, Human cervical cancer cell line Hela, human hepatic cancer cell line HepG 2 and human breast cancer cell line PMC with different relative molecular mass(Mr), but no expression of the protein ingredient in the normal human heart and fresh lung tissues. The Mr of lung cancer associated antigen N35 was 75×10 3—130×10 3 and was determined as aglycoprotein by N glycanase digestion with 3 fragments of Mr being 65×10 3,84×10 3 and 86×10 3. The differential centrifugation and subcellar assay showed the lung cancer associated antigen N35 to be distributed on the nuclei significantly much more than at the mitochondria and membrane. The result by immunoflorescence staining of the mitotic cells also showed that lung cancer associated antigen N35 was located on the centriole of the chromosomal domain at thd S G2 phage. Conclusions:Lung cancer associated antigen N35 might be a very important tumor associated antigen controlling the malignant proliferation of cancer cells.

关 键 词：单克隆抗体 肺癌 相关抗原 亚细胞结构 

分 类 号：R734.202[医药卫生—肿瘤]