Analysis of phosphotyroisine-containing proteins related to the malignancy in gliomas by proteomic differential display

胶质瘤是最常见的颅内原发肿瘤。利用临床和组织学评估的不同级别的胶质瘤，我们用双向凝胶电泳质谱方法研究了它们的磷蛋白谱，并鉴定了差异表达的磷蛋白，这些差异表达蛋白可能是了解这些进展的有用的分子指标。对同一患者的不同级别的胶质瘤标本或不同患者的不同级别的胶质瘤标本的磷蛋白图谱进行检测，发现约40种不同的、差异表达的磷蛋白在胶质瘤进展过程中表达。这些感兴趣的磷酸蛋白被挑选出来，凝胶内消化和质谱学指纹图谱。其中一些成功地进行了鉴定。这些磷蛋白属于多种功能基团，如细胞骨架和中间丝蛋白、热休克蛋白、酶和调节蛋白。一些在III级和多形性胶质母细胞瘤(GBM)中差异表达，而另一些与特定级别相关。此前已有报道称，有几种蛋白质在人脑癌中以非磷酸化形式差异表达。值得注意的是，在III级和GBM肿瘤中，HSPs和胶质纤维酸性蛋白(GFAP)以及胶质纤维酸性蛋白(GFAP)均被磷酸化。我们试图从它们的联合作用来解释胶质瘤的恶性和进展。与非肿瘤组织相比，白蛋白、过氧化还蛋白1、果糖二磷酸缩醛酶、肌酸激酶、hnRNPs、ATP依赖的DNA解旋酶在GBM组织中表达上调。这些差异表达的蛋白质为正常脑和胶质瘤之间存在的差异提供了新的信息，因此可能被证明是有用的诊断或预后价值的分子指标。

Gliomas are the most common of the primary intracranial tumors. Using clinically and histologically assessed various grade of gliomas, we have studied their phosphoprotein profiles by a two-dimensional gel electrophoresis-mass spectrometry approach and identified differentially expressed phosphoproteins which may be useful molecular indicators to understand these progression. Examination of the phosphoprotein profiles of glioma samples of different grades from same patient or different grades from different patients revealed about 40 distinct, differentially expressed phosphoproteins during glioma progression. These phosphoproteins of interest were picked, in-gel digested and mass spectrometry fingerprinted. Some of them were identified successfully. These phosphoproteins belonging to various functional groups such as cytoskeleton and intermediate filament proteins, heat shock proteins, enzymes and regulatory proteins. Some were found to be differentially expressed in both Grade III and glioblastoma multiform (GBM) while others were associated with a particular grade. Several proteins had been reported previously as differentially expressed in human brain cancer in non-phosphorylated form. A notable observation was that prohibitin, a potential tumor suppressor protein, and HSPs as well as glial fibrillary acidic protein (GFAP), major protein of the glial filaments were phosphorylated in both grade III and GBM tumors. We attempt to explain glioma malignancy and progression in terms of their combined role. Albumin, peroxiredoxin 1, aldolase C fructose-biphosphate, creatine kinase, hnRNPs, ATP-dependent DNA helicase were up regulated in GBM versus non-tumor tissues. These differentially expressed proteins provided novel information on the differences existing between normal brain and gliomas, and thus might prove to be useful molecular indicators of diagnostic or prognostic value.