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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。
Objective To analyze the relevant characteristics of lung cancer related protein N35 and to explore its potential clinical value. Methods Anti-human lung cancer mAbN 35 was used as an immunoprecipitation probe to detect the expression of its related antigen in lung cancer tissue, lung cancer cell line GLC 82, cervical cancer cell line HeLa, liver cancer cell line HepG 2, breast cancer cell line PMC, normal human heart and lung tissue; using N-glycan enzymatic hydrolysis method to determine the relationship between lung cancer-associated protein N35 and glycoprotein molecules; using differential centrifugation technique to isolate the lung adenocarcinoma cell line GLC 82 subcellular The distribution of cell membrane, nucleus and mitochondria was detected by immunoblotting. The distribution of N35 in subcellular structure was detected by immunoblotting. The functional structure of lung cancer associated protein N35 in mitosis was detected by immunofluorescence staining. Results The lung cancer related protein N35 was a kind of glycoprotein which was not found in the normal human heart and lung tissue protein components but distributed in GLC 82, HeLa, HepG 2 and PMC cell proteins with different molecular weight. Subcellular structure is mainly distributed in the nucleus, followed by the mitochondria, the membrane distribution on the least, does not support its presence as a membrane protein or transmembrane protein; in tumor cell mitosis into S to G2 period, clearly located in the center Centriole structure, suggesting that its function may be related to the unlimited proliferation of tumor cells. Conclusions N35, a lung cancer-related protein, is a protein that exists only in tumor cells and is closely related to its proliferation. It may play a role in regulating tumor cell growth.