【作者】 商崇智；

【导师】 张赛；

【作者基本信息】 天津医科大学 ， 外科学， 2009， 硕士

【摘要】 背景：胶质瘤是最常见的颅内恶性肿瘤,病情进展快,早期多无明显临床表现,大部分患者就诊时已是中晚期,由于本身具有侵袭性生长的特性,与正常脑组织无明显界限,多数不限于一个脑叶,手术常无法全切,即使辅助以放、化疗,其疗效仍较差,复发率和死亡率仍较高,严重危害人类健康。早期发现和鉴别胶质瘤对胶质瘤的治疗意义重大。随着影像学的发展,胶质瘤的临床诊断取得了长足的进步,但是胶质瘤的生物学诊断仍缺乏有效的手段,造成这一现状的主要原因是胶质瘤缺乏敏感的、特异的生物学标志物。近年来蛋白组学的飞速发展,给寻找新的脑胶质瘤标志物提供了新的技术平台。而早期诊断与鉴别有赖于对胶质瘤标本、血浆、脑脊液中特异肿瘤生物学标志物的发现。胶质瘤血清蛋白表达谱的研究国内外很少,可能是由于血-脑屏障导致胶质瘤特异性蛋白在血清表达中很少。脑胶质瘤细胞的代谢产物可直接进入脑脊液,因此脑脊液中蛋白质的特异性改变可动态反映脑胶质瘤的代谢状态,因此对脑脊液中蛋白质组分进行分析可能找到胶质瘤特征性生物标志物。然而胶质瘤生物学标记研究中应用最多的还是对肿瘤标本或细胞系进行分析,以寻找与肿瘤诊断、良恶性区分的肿瘤标志物。本文对恶性胶质瘤细胞系进行蛋白质组学研究,希望尽可能多地寻找到参与国人胶质瘤恶性演变的特异蛋白质,将有助于进一步阐明胶质瘤发病机理、寻找治疗靶点、进行临床用药筛选和新药物的研发。目的：建立国人恶性胶质瘤细胞系的蛋白质组差异表达谱,为以其为基础的相关研究提供必要参考。尽可能多地寻找到参与国人胶质瘤恶性演变的关键蛋白质,对于阐明胶质瘤发病机理、寻找治疗靶点、进行临床用药筛选和新药物研发等提供理论依据和数据参考。方法：提取3例国人恶性胶质瘤细胞系CHG-5、TJ899和TJ905的总蛋白,等比例混匀后行双向电泳,正常胶质细胞为平行对照,筛选和鉴定出差异蛋白质点,并就其中部分蛋白质进行细胞免疫化学方法验证。结果：共筛选出13个差异蛋白质点,5个蛋白表达下降,8个表达水平上升,其中10个得到成功鉴定,它们主要分属于细胞骨架蛋白、代谢相关蛋白、肿瘤细胞迁移,以及应激与炎症反应相关蛋白。细胞免疫化学检测结果与蛋白质组技术结果相似。结论：国人恶性胶质瘤细胞系与正常胶质细胞比较具有明显的多个差异蛋白表达,它们可能共同参与脑胶质细胞的瘤变过程。更多还原

【Abstract】 Background: Glioma is the most commonest intracranial malignant tumor. Glioma cells proliferate rapidly.lt has no obvious clinical symptoms in early stage. So many patients seek medical treatment till late stage.Because of its character of invasive growth, no obvious boundaries between normal brain tissues, the majority is not limited to one brain lobe, surgery often can not totally resect tumor tissues, even if combining radiotherapy and chemotherapy, its clinical effect is still poor, the recurrence rate and mortality is still high,it seriously endanger human health. Early detection and identification of glioma has great significance for glioma therapy. With the development of imaging, clinical diagnosis of glioma has made considerable progress, but the biological diagnosis of glioma is still lack of effective means, resulting in this situation is mainly due to lack of sensitive, specific biological markers. Proteomics has developed rapidly in recent years, and it will provide a new technology platform for finding many new tumor biological markers. And early diagnosis and identification depend on specific tumor biomarker discoveries of glioma specimens, plasma, cerebrospinal fluid. Human serum protein expression profiling study is rare at home and abroad, may be due to the blood-brain barrier leading to specific tumor protein expression in serum is very slow.Glioma cell metabolites secrete directly into the cerebrospinal fluid, so the specific protein in cerebrospinal fluid reflect glioma metabolism changes dynamically, the protein component analysis of cerebrospinal fluid may find a specific tumor markers. However glioma biomarker research is still most wide using tumor specimens or cell lines for analysis to find tumor markers for diagnosis, distinguishing benign or malignant tumor.We conducted proteomics research on malignant glioma cell lines in order to find much more specific proteins which participate in the malignant evolution of glioma, it will be helpful to further elucidating the pathogenesis, looking for therapeutic targets, screening clinical drug and developing new drugs.Objective:To establish the differential proteomics expression profiles of malignant glioma cell lines from Chinese, and provide reference for other basic related researches. To find key proteins as much as possible which participate in the malignant evolution of glioma from Chinese, and provide theoretical basis and reference data for elucidating the pathogenesis of gliomas, looking for therapeutic targets, screening clinical drugs and developing new drugs.Methods: The total protein was extracted from 3 gliobma cell lines, named CHG-5, TJ899 and TJ905. After an even mixing, the total protein was presented by two-dimensional (2D) electrophoresis, scanned and analyzed. Part of the identified protein spots were verified by immunocytochemistry. The glia cells were used as control through the study.Results:13 differential protein spots were selected,5 protein expression level decreased, eight protein expression level increased,and 10 protein spots were identified successfully, which probably involved in cytoskeleton forming, cellular metabolism, tumor migration, stress and inflammatory reaction. These results were coincident with that from immunocytochemistry.Conclusion:There were obviously differential proteomics expression between glioma cell lines and normal glia cells, and they may play key roles in the transform progress of glia to glioma.更多还原