肿瘤相关蛋白亚细胞结构定位的研究

The Subcellular Localization of Tumor Associated Protein

目的应用激光共聚焦显微镜技术对肺癌相关蛋白N35进行亚细胞定位,观察它们在肿瘤细胞中的分布和肿瘤细胞周期不同时期的变化,并与其它组织和肿瘤进行比较,了解其特异性。方法利用抗人肺癌单克隆抗体N-35作为免疫探针作免疫荧光标记,应用激光共聚焦技术观察肿瘤相关蛋白亚细胞结构的定位,测定其相关抗原在肺癌细胞系GLC—82、人淋巴细胞系、人胚肺二倍体细胞系(KMB—17)、人皮肤纤维细胞系的存在及分布情况以及在肿瘤细胞周期不同时期的变化。结果肿瘤相关蛋白N35不存在于正常人淋巴细胞、正常肺细胞及人皮肤纤维细胞中,而存在于肺癌细胞蛋白中,在亚细胞结构中主要分布于胞核,在肿瘤细胞有丝分裂进入G2至M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,其功能可能与肿瘤细胞无限制增殖活动有关,是肿瘤标志物之一。可用于临床肿瘤的早期诊断、疗效评估、监测复发、预测转移和推测预后。

Objective The study of the lung cancer associaced protein N35 were located in subcellular structers by laser scanning confocal microscope. Their distribution in tumor cell and their change in different stages of tumor cell cycle were watched, to identify and characterize it and to learn its potential clinical application. Methods cells culture, ELISA, immunof lourescence marking and laser scanning confocal microscope have been used by the monoclonal antibody N-35 as the immunoprdoe to detect the activity of N35, the subcellular localization of tumor associated protein, the distribution of lung cancer associated antigen N35 among lung cancer cell lines and normal human tissue, change of different stages in tumor cell cycle .Results Tumor associated protein N35 distributed in the lung cancer cell live GLC-82, but no expresstion of the protein ingredient in the normal human lymphocyce, the normal human lung cell, human skin fibroblasts, distributed on the nuclei significantly. Tumor associated protein N35 was located on the centriole of the chromosomal domain at the S-G2 phage.Conclusions The tumor associated protein N35 might be a tumor cell growth regulator, expressed only it＇ s the cancer cells and related with the proliferation of cancer cells. It is one of tumor markers. It could be used to diagnose clinical tumor earlier, evalue therapy effect, monitor recurrence, predict metastasis and speculate prognosis.