N35肺癌相关蛋白的特性  

Characterization of tumor associated protein of N35 lung cancer

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：《癌症》2000年第9期859-863,共5页Chinese Journal of Cancer

基　　金：本课题受国家自然科学基金!(编号39960078)资助

摘　　要：目的：分析和确定N35肺癌相关蛋白的有关特性及其潜在的临床应用价值。方法：利用抗人肺癌单克隆抗体N35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GLC82、宫颈癌细胞系Hela、肝癌细胞系HepG2、乳腺癌细胞系PMC、正常人心脏及肺组织中的存在及分布情况；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；采用差速离心技术分离出肺腺癌细胞系GLC82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蛋白N35在亚细胞结构中的分布状况并与免疫组化结果相印证；用免疫荧光技术探测肿瘤相关蛋白N35在肿瘤细胞有丝分裂过程中的染色体结构定位；用多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：肿瘤相关蛋白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，而以不同分子量形式分布于GLC82、Hela、HepG2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，似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，强烈提示其功能可能与肿瘤细胞无限制增殖活动有关；麦芽凝集素亲和层析法是大量纯化肿瘤相关蛋白N35最有效的途径。结?Objectives： To identify and characterize N35 lung cancer associated protein and to learn its potential clinical application. Methods： Immunoprecipitation, immunoblotting, differential centrifugation and subcellular assay, immumnohistochemistry,N glycanase digestion, mitotic cell immunoflourescence and multiple methods of affinity chroma tography have been applied by using monoclonal antibody N 35,to detect the distribution of the protein N35 among various cancer cell lines and normal human tissues, the relationship between the protein N35 and glycoprotein,the location of the subcellular structure and chromosomal domain of the protein N35, and the most effective way of purification of tumor associated protein N35 accordingly. Results： The protein N35, a glycoprotein, distributed with different molecular size in human 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 molecular weight, but not in normal human heart and lung fresh tissue. The protein pressented mainly at the nucleu, less at the mitochondrail and membrane, and located at the centriole of the chromosomal domain when the cell entered S through G2 phase. Conclusion： The tumor associated protein N35 might be a tumor cell growth regulator, expressed only in the cancer cells and related with the proliferation of cancer cells .

关 键 词：肺肿瘤 肿瘤相关蛋白 糖蛋白分子 N35 

分 类 号：R734.2[医药卫生—肿瘤]