Pygo2在脑胶质瘤组织和细胞的表达及临床意义

目的:检测Pygo2在脑胶质瘤组织和细胞中的表达,并探讨其临床意义.方法:采用RT-PCR检测5种胶质瘤细胞株Pygo2 mRNA表达,Real-timePCR检测67例Ⅱ～Ⅳ级脑胶质瘤组织中Pygo2 mRNA表达,免疫组织化学SP法检测80例脑胶质瘤组织中Pygo2的蛋白表达.结果:Pygo2 mRNA在5种胶质瘤细胞株中均有表达,其中U251细胞中最高,C6细胞中最低.Pygo2mRNA在正常脑组织中表达极低,而在脑胶质瘤Ⅱ～Ⅳ级表达显著增高,且Pygo2mRNA表达随肿瘤分级的增高而显著增强(P＜0.05).Pygo2在脑胶质瘤组织细胞中定位于胞核,在正常脑组织中表达呈阴性,而在脑胶质瘤组织中约有65.0%阳性表达,两者差异显著(P＜0.01).Pygo2表达Ⅱ级(WHOⅡ)中低,Ⅳ级(WHO Ⅳ)中高.Pygo2在脑胶质瘤Ⅰ级、Ⅱ级、Ⅲ级、Ⅳ级中表达阳性率分别是5/13(38.5%)、12/26(46.2%)、15/19(78.9%)、20/22(90.9%),Pygo2蛋白表达与肿瘤分级密切相关(P＜0.01).此外,Pygo2蛋白表达水平与患者的性别、年龄均无显著性差异(P＞0.05).结论:Pygo2在正常脑组织中呈阴性,在脑胶质瘤组织和细胞中均高表达,且在组织中其表达随胶质瘤级别增高而显著增强,提示Pygo2可能在脑胶质瘤的发生、恶性演进中起了重要作用.     

AR、PCNA表达与脑膜瘤临床病理因素的相关性研究

目的探讨雄激素受体(AR)在脑膜瘤组织中的表达及其与肿瘤增殖潜力的关系.方法采用免疫组化SABC法,检测AR、增殖细胞核抗原(PCNA)在39例脑膜瘤组织中的表达.结果51.3%(20/39)的脑膜瘤组织中AR呈不同程度的表达.良性、非典型性、恶性脑膜瘤中AR表达率分别为31.6%(6/19)、58.3%(7/12)、87.5%(7/8).恶性脑膜瘤中AR阳性细胞数显著高于非典型性和良性脑膜瘤(P＜0.05).恶性脑膜瘤平均PCNA标记指数(PCNA U)明显高于非典型性(P＜0.05)和良性脑膜瘤(P＜0.05).AR阳性脑膜瘤的PCNA U高于AR阴性脑膜瘤(P＜0.05),AR表达与PCNA的表达呈正相关.结论脑膜瘤AR表达与其病理级别有关,AR参与了肿瘤的生长和血管生成;检测脑膜瘤中AR表达可间接反映肿瘤的增殖潜力.     

核因子-κB在人脑肿瘤中的表达及意义

目的 :探讨核因子 κB(nuclearfactorkappaB ,NF κB)在人脑肿瘤组织中的表达及意义。方法 :应用免疫组织化学方法检测NF κBP65在 1 0 6例人脑肿瘤中的表达 ,其中星形细胞肿瘤 64例、髓母细胞瘤 2 2例、脑膜瘤 2 0例。 1 0例正常大脑组织作为对照。结果 :星形细胞肿瘤、髓母细胞瘤及脑膜瘤中NF κBP65蛋白阳性表达率分别为54 7% (35 64)、36 4% (8 2 2 )和 5 0 % (1 2 0 ) ;1 0例正常脑组织中NF κBP65无 1例阳性表达。NF κBP65蛋白的表达在星形细胞瘤与脑膜瘤、髓母细胞瘤与脑膜瘤之间差异显著 (P <0 0 5)。结论 :NF κB组成性活化与脑恶性肿瘤的发生相关 ,NF κBP65是一种与人脑肿瘤相关的新的癌蛋白     

5-脂氧合酶及Ki-67在星形细胞瘤中的表达及其相关性研究

目的:研究5-脂氧合酶(5-LOX)和Ki-67在星形细胞瘤中的表达及相关性.方法: 免疫组织化学法检测60例星形细胞瘤(低度恶性33例,高度恶性27例)中5-LOX和Ki-67的表达,逆转录聚合酶链反应(RT-PCR)方法检测5-LOX和Ki-67的mRNA水平,另取14例行脑外伤内减压术的脑组织作为对照.结果: 对照组中5-LOX和Ki-67不表达或低表达,星形细胞瘤中5-LOX阳性表达率66.7%(40/60),Ki-67阳性表达率71.7%(43/60);高度恶性组5-LOX和Ki-67 的阳性表达率高于低度恶性组(P<0.01);5-LOX和Ki-67表达呈正相关(γ＝0.875,P<0.01).星形细胞瘤5-LOX和Ki-67的mRNA含量高于正常脑组织(P<0.01),高度恶性组5-LOX和Ki-67的mRNA含量高于低度恶性组(P<0.01),5-LOX和Ki-67在星形细胞瘤中的mRNA水平呈正相关(γ＝0.781,P<0.01).结论: 5-LOX和Ki-67的表达与星形细胞瘤恶性程度有关,二者在星形细胞瘤发生发展过程中有协同作用.     

NF-κB与PCNA在人脑星形细胞瘤中的表达

目的:探讨NF-κB与PCNA在人脑星形细胞瘤以及正常脑组织中的表达情况及其与病理分级之间的相互关系.方法:SP免疫组织化学方法检测77例不同级别的星形细胞瘤及正常脑组织标本NF-κB p65与PCNA的表达情况.结果:除了在星形细胞瘤Ⅲ、Ⅳ级之间无显著性差异外(P＞0.05),PCNA在其余各级别星形细胞瘤中的表达有显著性差异(P＜0.05),其阳性率随着肿瘤级别的增高而升高.尽管NF-κB的表达总体差异有显著性(P＜0.01),但NF-κB在星形细胞瘤相邻各级别之间的表达无显著性差异(P＞0.05),然而高度恶性组(Ⅲ～Ⅳ级)阳性率显著高于低度恶性组(Ⅰ～Ⅱ级)阳性率,二者有显著性差异(P＜0.01).结论:NF-κB异常表达可能通过PCNA影响肿瘤细胞增殖活性,免疫组织化学方法联合检测NF-κB与PCNA的表达情况能客观反映星形细胞瘤的增殖恶性程度及预后.     

热休克蛋白70对星形细胞瘤凋亡调控的研究

目的 :研究热休克蛋白 70 (HSP70 )在星形细胞瘤 (astrocytoma)中的表达与意义。方法 :采用鼠抗人 HSP70单克隆抗体及免疫组织化学 SABC法。结果 :在 99例星形细胞瘤中 ,HSP70表达 70例 (70 .7% ) ;用于对照的正常脑组织中 ,HSP70阳性仅 1例 (10 % )。HSP70在星形细胞瘤中的表达强于正常对照组 (P<0 .0 5) ,HSP70在 级和 级、 级和 级、 级和 级星形细胞瘤相互之间表达无显著性差异 (P>0 .0 5) ,但在 50例低度恶性星形细胞瘤 (WHO 级～ 级 )中表达强于在49例高度恶性星形细胞瘤 (WHO 级～ 级 )中的表达 (P<0 .0 5)。结论 :HSP70在大部分星形细胞瘤中有不同程度表达 ,HSP70可能参与星形细胞瘤的凋亡调变 ;其表达与星形细胞瘤分化有关     

中期因子蛋白在脑星形细胞瘤中的表达

[目的]探讨中期因子(MK)蛋白在脑星形细胞瘤中的表达及其与微血管密度(MVD)、组织分级和预后的关系.[方法]应用免疫组织化学方法检测52例星形细胞瘤组织中MK蛋白的表达和MVD计数.[结果]52例星形细胞瘤组织中MK蛋白阳性率为63.5%(33/52),MK阳性染色定位在瘤细胞胞质内.其中Ⅰ～Ⅱ级和Ⅲ～Ⅳ级阳性率分别为44.4%、73.5%,两者间有显著差异(P＜0.05);星形细胞瘤组织中MVD均值为71.8±27.8,其中,Ⅰ～Ⅱ级和Ⅲ～Ⅳ级MVD分别为52.8±18.4、81.7±26.9,两者间差异有显著性(P＜0.01);星形细胞瘤MK蛋白阳性组MVD高于阴性组(P＜0.01).术后生存期＜2年的患者MK蛋白阳性率及MVD均高于术后生存期≥2年者(P＜0.05,P＜0.01).[结论]MK的蛋白表达水平能反映星形细胞瘤的恶性程度,可作为星形细胞瘤预后分析的指标.     

人脑胶质瘤中CD147的表达及其意义

背景与目的:以往研究发现CD147与某些恶性肿瘤存在相关性。本研究探讨CD147在人脑胶质瘤中的表达及其与病理分级的相关性。方法:收集61例星形细胞肿瘤及18例瘤旁脑组织标本,制作组织芯片。进行免疫组织化学染色,用半定量逆转录聚合酶链反应法(RT-PCR)进行CD147mRNA检测。结果:(1)胶质瘤组织中CD147蛋白的阳性表达率为74.3%,正常脑组织中仅血管内皮细胞浆有阳性着色。(2)III、IV级星形细胞肿瘤瘤细胞胞浆CD147着色显著强于I、II级低级别星形细胞肿瘤。(3)CD147mRNA在胶质瘤组织中阳性率为83.3%,明显高于正常脑组织中的阳性率25%(P<0.05)。CD147mRNA在星形细胞肿瘤组织中的表达量为0.274±0.087,明显高于正常组织0.081±0.043,两者差异具有显著性(P<0.05)。结论:CD147的表达可能在人脑胶质瘤的发生发展中起重要作用。     

EMP1基因在人脑胶质瘤中的表达及其临床意义

目的:探讨上皮膜蛋白1基因(epithelial membrane protein 1,EMP1)在人脑胶质瘤中的表达及其与肿瘤恶性程度的相互关系。方法:肿瘤组织标本均取自上海长征医院神经外科2005至2006年部分手术病例,正常脑组织来源于捐献,分别采用逆转录酶聚合酶链反应(RT-PCR)和免疫组化染色方法检测33例星形细胞瘤、3例少枝胶质细胞瘤、2例室管膜瘤以及7例正常脑组织标本中EMP1 mRNA及蛋白的表达。结果:EMP1无论在基因还是在蛋白表达水平上,在胶质瘤和正常脑组织中均有不同程度表达,但在星形细胞瘤中表达的含量明显高于正常脑组织,且在星形细胞瘤中含量随着其病理分级的增高而增高,在低级别(WHOⅠ～Ⅱ级)与高级别(Ⅲ～Ⅳ级)之间存在显著差异(蛋白P<0.05;基因P<0.01)。结论:基因EMP1在人脑胶质瘤中显著高表达,并且与星形细胞瘤的恶性程度密切相关。     

脑膜瘤端粒酶的活性研究

目的探讨端粒酶作为脑膜瘤标志物,在肿瘤良恶性鉴别、恶性程度评估以及预后预测等方面的意义.方法利用TRAP-ELISA方法定量检测14例脑膜瘤中端粒酶的活性,结合肿瘤的分类和患者的临床资料分析.另取正常脑组织标本10例作为对照.结果10例脑膜瘤中,1例复发者表现为端粒酶阳性(10%),4例恶性脑膜瘤中3例呈端粒酶阳性(75%),两组之间比较差异有显著性(P=0.041),另外1例脑膜肉瘤呈端粒酶阳性.10例正常脑组织中未检测到端粒酶活性.结论端粒酶活性的激活在恶性脑膜瘤中是常见的现象.端粒酶作为肿瘤标志物,在脑膜瘤的良恶性鉴别、恶性程度评估、预后预测等方面具有重要的意义.     

Molecular characterization of a novel human brain tumor-associated gene BR-3

Using the technique of differential hybridization of a human fetal brain library, we have identified a novel gene, brain 3 (BR-3). This gene is not expressed in normal human brain tissue samples but is expressed at high levels in human low-grade glioma tissue samples. We have cloned and sequenced the full-length cDNA corresponding to this gene. Data base search analysis indicated that the BR-3 gene has strong homology to a genomic sequence present on chromosome 1 but no homology to expressed genes. Open reading frame analysis has indicated the presence of a 71 amino acids long protein sequence. A data base search for the protein sequence homology showed no similarity to known sequences. Expression analysis of BR-3 indicated that it is expressed at high level in six out of seven low-grade glioma samples analyzed. In addition low levels of BR-3 gene expression was observed in six out of seven anaplastic astrocytoma tissue samples analyzed. BR-3 expression was observed in four of eight glioblastoma samples analyzed. Expression analysis of normal human tissues indicated that it is expressed in kidney, skeletal muscle, lung, spleen and heart. No expression of the BR-3 gene was observed in brain, liver or testes tissue. To understand the role of the BR-3 gene in cancer in general, we studied its expression in other cancer cell lines. Except for lung and ovarian carcinoma, the BR-3 gene is expressed in breast carcinoma, colon adenocarcinoma, prostatic adenocarcinoma, and pancreatic adenocarcinoma tissue samples. On the basis of its sequence, unique expression pattern, we conclude that BR-3 gene product may play a critical role in the genesis of human gliomas tumors.     

Expression of tissue factor pathway inhibitor 2 inversely correlates during the progression of human gliomas.

Protease inhibitors regulate a variety of physiological and pathological processes including angiogenesis, embryo implantation, intravascular fibrinolysis, wound healing, and tumor invasion. Tissue factor pathway inhibitor (TFPI) 2 is a Mr 32,000 Kunitz-type serine protease inhibitor that inhibits plasmin, trypsin, chymotrypsin, cathepsin G, and plasma kallikrein but not urokinase-type plasminogen activator, tissue plasminogen activator, or thrombin. In this study, we determined the relative amounts of TFPI-2 in low-, intermediate-, and high-grade human glioma cell lines and tumor tissue samples. TFPI-2 protein and mRNA levels (measured by Western and Northern blotting) were highest in low-grade glioma cells (Hs683), lower in anaplastic astrocytoma cells (SW1088 and SW1783), and undetectable in high-grade glioma cells (SNB19). Analysis of TFPI-2 protein in human normal brain and in glioma tumor tissues for TFPI-2 revealed the highest levels in normal brain, lesser amounts in low-grade gliomas and anaplastic astrocytomas, and undetectable amounts in glioblastomas. In situ hybridization of TFPI-2 mRNA with normal brain tissues revealed the greatest positivity in neurons, with moderate positivity in both glial and endothelial cells and moderate, little, or no TFPI-2 mRNA in low-grade glioma, anaplastic astrocytoma, and glioblastoma tumor tissue samples, respectively. We also found that recombinant TFPI-2 inhibited the invasiveness of SNB19 glioblastoma cells in a Matrigel assay in a dose-dependent manner. Collectively, these results suggest that TFPI-2 has a regulatory role in the invasiveness of gliomas in vitro and in vivo.     

Expression of emmprin (CD147), a cell surface inducer of matrix metalloproteinases, in normal human brain and gliomas

EMMPRIN (extracellular matrix metalloproteinase inducer), also called CD147, basigin or M6 in the human, is a member of the immunoglobulin superfamily that is present on the surface of tumor cells and stimulates adjacent fibroblasts to produce matrix metalloproteinases (MMPs). In our study, we investigated expression of EMMPRIN in human normal brain and gliomas, since mouse basigin and chicken HT7, the species homologues of human EMMPRIN, are associated with neuronal interactions and normal blood-brain barrier function, respectively. EMMPRIN expression was detected in all samples of non-neoplastic brain and glioma tissues examined. However, expression levels of EMMPRIN mRNA and protein were significantly higher in gliomas than in non-neoplastic brain. Moreover, levels of mRNA expression and immunohistochemical staining correlated with tumor progression in gliomas: They were highest in the most malignant form of glioma, glioblastoma multiforme, followed by anaplastic astrocytoma and then low-grade astrocytoma. Also, immunolocalization revealed quite different distributions in non-neoplastic brain and glioma: EMMPRIN was demonstrated only in vascular endothelium in non-neoplastic regions of the brain, whereas it was present in tumor cells but not in proliferating blood vessels in malignant gliomas. These data indicate that an MMP inducer molecule EMMPRIN is differently expressed in human normal brain and gliomas and could be associated with astrocytoma progression. Possible mechanisms whereby glioma cell EMMPRIN could influence tumor progression will be discussed.     

Reactivation of insulin-like growth factor binding protein 2 expression in glioblastoma multiforme: a revelation by parallel gene expression profiling.

We carried out a gene expression profiling study using cDNA array technology with 24 primary glioma tissues of low-grade (oligodendroglioma), intermediate-grade (anaplastic oligodendroglioma and anaplastic astrocytoma), and high-grade (glioblastomas multiforme) tumors and found that insulin-like growth factor binding protein 2 (IGFBP2) was consistently overexpressed only in glioblastoma multiforme. The cDNA array results were confirmed by Northern and Western blotting. The fact that the IGFBP2 gene, which is normally expressed in fetal cells and turned off in adult cells, becomes reactivated in the most advanced stage of glioma suggests that glioma progression is a result of dedifferentiation or results from a block of differentiation. Identification of IGFBP2 as a gene associated with glioma progression demonstrates the power and utility of high-throughput gene expression profiling in cancer gene discovery.     

Tumor Grade versus Expression of Invasion-Related Molecules in Astrocytoma

Peritumoral infiltration is characteristic of astrocytomas even in low-grade tumors. Tumor cells migrate to neighbouring tissue and cause recurrence. The extracellular matrix (ECM) plays a role in tumor invasion; expression levels of its components’ have been linked to tumor invasion. This study determines the mRNA and protein expression of 20 invasion-related ECM components by examining non-tumor brain; grade I-II-III astrocytoma and glioblastoma samples. Expression levels were measured by QRT-PCR and mass-spectroscopy. The connection between the expression pattern and tumor grade is statistically analyzed. During the analysis of data, key molecules (brevican, cadherin-12, fibronectin and integrin-β1) correlating the most with tumor grade were selected. While the mRNA level of brevican, ErbB2, fibronectin, integrin-β1 and versican discriminates low-grade from high-grade gliomas, of proteins RHAMM, integrin-α1 and MMP2 seems important. The expression pattern was found to be distinctive for tumor grade, as statistical classifiers are capable of identifying an unknown sample’s grade using them. Furthermore, normal brain and glioma expression patterns, along with low-grade astrocytoma and glioblastoma samples, differ the most. Determining the invasion-related molecules’ expression profile provides extra information regarding the tumor’s clinical behavior. Additionally, identifying molecules playing a key role in glioma invasion could uncover potential therapeutic targets in the future.     

应用组织芯片技术研究HDAC6在胶质瘤组织中的表达及预后意义

目的:探讨胶质瘤组织中HDAC6的表达及其与临床病理资料之间的关系.方法:采用免疫组化法检测胶质瘤组织芯片(正常脑组织20例、癌旁组织27例、胶质瘤组织206例)中HDAC6的表达量及其与临床病理资料之间的关系,应用SPSS 16.0软件分析胶质瘤组织中HDAC6表达水平与胶质瘤临床病理之间的相关性及其对胶质瘤患者预后的影响.结果:HDAC6在胶质瘤中的表达量明显高于正常脑组织和癌旁组织(P<0.05),并且在高级别胶质瘤(III-IV)中的蛋白表达量明显高于低级别胶质瘤(I-II)中的蛋白表达量.依据生存分析,发现HDAC6蛋白在低表达组中与高表达组相比有生存优势,PFS(无进展生存期)及OS(总生存期)均延长,差异具有统计学意义(P<0.05).结论:HDAC6在胶质瘤中的表达量明显升高,促进了肿瘤的发生和发展,通过HDAC6表达水平可预测胶质瘤患者的预后.     

Brevican,Neurocan, Tenascin-C and Versican are Mainly Responsible for the Invasiveness of Low-Grade Astrocytoma

The extent of tumor removal determines the effectiveness of postoperative oncotherapy. This is especially true for primary brain tumors, where peritumoral invasion usually makes radical resection impossible. The aim of the study was to determinate the specific expression pattern of invasion related molecules of different intracranial tumors and to identify molecules that are principally responsible for the peritumoral invasiveness of grade II astrocytoma mRNA expression of 26 extracellular matrix (ECM) molecules was determined in tissue samples from grade II astrocytoma, schwannoma, intracerebral metastases of non-small cell lung cancer and normal brain. Immunohistochemical staining for brevican, neurocan, tenascin-C and versican was also performed for each tumor group. Comparing astrocytoma to metastasis, schwannoma and normal brain; and metastasis and schwannoma to normal brain, 22, 17, 20, 21, and 19 molecules, respectively, were found to be significantly overexpressed at the mRNA level. Cluster analysis of mRNA expression showed a specific gene expression pattern for each histological group. Four molecules of 26 were found to be associated to astrocytoma. Immunohistochemical staining confirmed the results of the mRNA analysis at the protein level. Tumors of different origin have a specific invasive phenotype that can evidently determinate on gene expression level. This characteristic expression pattern of the invasion-related molecules might help to screen exact targets for anti-invasion drugs. In case of low-grade astrocytoma. brevican, neurocan, tenascin-C and versican were found to correlate principally with the invasive phenotype of low-grade astrocytoma, thus these molecules can potentially serve as targets for anti-invasion therapy in the future.