肺癌细胞相关蛋白分子N35的实验研究  

Experimental study on associated protein mol-ecule N35 in human lung cancer cells

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：《细胞与分子免疫学杂志》2000年第2期152-155,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金!资助 ;No.39960078;云南省九五重点学科肿瘤学经费资助

摘　　要：目的分析肺癌相关蛋白N35的有关特性 ,探讨其潜在的临床应用价值。方法以抗人肺癌mAbN 35作为免疫探针 ,用免疫沉淀和免疫印迹法 ,测定其相关抗原在肺癌组织、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与分布情况 ;用N 聚糖酶酶解方法确定肺癌相关蛋白N35与糖蛋白分子的关系 ;用差速离心技术分离出肺腺癌细胞系GLC 82亚细胞结构中的胞膜、胞核及线粒体成分 ,经免疫印迹法测定相关蛋白N35在亚细胞结构中的分布状况 ;用免疫荧光技术检测肺癌相关蛋白N35在肿瘤细胞有丝分裂过程中的功能结构定位。结果肺癌相关蛋白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 ,而是以不同分子质量的形式分布于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 ,不支持其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 ,提示其功能可能与肿瘤细胞的无限制增殖有关。结论肺癌相关蛋白N35是一种只存在于肿瘤细胞并与其无限增殖密切相关的蛋白分子 ,有可能具有调节肿瘤细胞生长的作用。Aim To identify and characterize the associated prolein of lung cancer and to evaluate its prospective possibility for the clinical application. Methods Immunoprecipitation, immunoblotting, differential centrifugation and subcellular assay, N glycanase digestion, mitotic cell immunoflourescence have been employed by using the monoclonal antibody N35 as the immunoprobe to detect the distribution of the prolein N35 among various cancer cell lines and normal human tissues, to determine the relationship between the protein N35 and glycoprotein, and to examine the location of the subcellular structure and chromosomal domain of the protein N35. Results The protein N35 is a glycoprotein, distributing 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), with no expression of the protein ingredient in the normal human heart and lung fresh tissues. The Mr of the associated protein N35 was 75×103-130×10 3 and was determined as a glycoprotein by N glycanase digestion with 3 fragments of Mr being 65×103,84×103 and 86×103. The differential centrifugation and subcelullar assay showed the higher distribution of associated protein N35 in the nuclei than in the mitochondrail and membrane. Immunoflouresccnce staining of the mitotic cells indicated that associated protein N35 was located at the centriole of the chromosomal domain at the S-G2 phage. Couclusion The associated protein N35 might be expressed only by the cancer cells and related with the proliferation of cancer cells as a role of tumor cell growth regulator.

关 键 词：肺癌细胞 肺癌相关蛋白N35 中心粒 

分 类 号：R734.2[医药卫生—肿瘤]