尿激酶,尿激酶受体在人脑胶质瘤中的免疫组织化学研究

研究尿激酶(uPA)、尿激酶受体(uPAR)在人脑胶质瘤中的表达特征,评估其在胶质瘤恶性生长方式中的作用。选取57例不同恶性程度的星形细胞瘤、13例良性脑膜瘤和10例正常脑组织,分别以抗uPA单克隆抗体和抗uPAR多克隆抗体用ABC法进行免疫组织化学染色和半定量分析。结果表明:uPA、uPAR的表达程度与胶质瘤的恶性程度有关,胶质瘤恶性程度越高,表达程度越高。良性脑膜瘤的表达程度较低。正常脑组织则未见uPA、uPAR的表达。uPA染色除见于肿瘤细胞、血管内皮细胞外,还见于少数胶质增生带中的正常胶质细胞和神经元。在肿瘤细胞增殖密集区域,uPA阳性的肿瘤细胞比例较低,而在肿瘤细胞生长较稀疏、肿瘤血管增殖旺盛的区域阳性细胞比例较高。uPAR的染色定位于肿瘤细胞和血管内皮细胞。uPA和uPAR的共同表达是浸润性生长的胶质瘤的特征之一。其表达程度可作为判断星形细胞瘤浸润性的重要参考指标。在人体内,表达uPA的细胞种类是较多的。uPA、uPAR的表达对胶质瘤的浸润和血管发生起着重要作用。     

纤溶酶原激活抑制因子—1与脑瘤恶性程度和坏死之间的关系

作者对197例脑瘤标本用酶联免疫吸附法测定纤溶酶原激活抑制因子—1(PAI—1)含量。其中恶性胶质瘤45例,转移瘤45例,肿瘤坏死区6例,脑膜瘤47例,低恶性胶质瘤26例,听神经瘤18例,正常脑组织10例。 结果发现正常脑组织PAI—1含量很低。恶性胶质瘤和转移瘤的PAI—1含量比低恶性胶质瘤和脑膜瘤明显增高。肿瘤坏死区PAI—1含量比恶性胶质瘤和转移瘤更高。恶性肿瘤PAI—1含量比良性肿瘤高3.5倍。统计学差异明显(P=0.0004)脑水     

葡萄糖转运蛋白1型在脑肿瘤中的表达及意义

目的探讨葡萄糖转运蛋白1型(Glucose transporter-1,GLUT1)在脑肿瘤中的表达及其意义。方法 收集55例颅脑手术标本,包括46例脑肿瘤患者和9例正常脑组织,采用单克隆抗体的GLUT1免疫组织化学SP法进行染色。结果 7例低度恶性胶质瘤中的3例和全部13例高度恶性胶质瘤不同程度的表达GLUT1;10例良性脑膜瘤中的3例和全部5例恶性脑膜瘤为阳性表达;6例垂体瘤中的3例和5例表皮样囊肿也异常表达;所有正常脑组织均未见表达。结论 四种脑肿瘤均可不同程度表达GLUT1,尤其在恶性肿瘤为明显,其与脑肿瘤的恶性程度相关。     

葡萄糖转运蛋白1型在脑肿瘤中的表达及意义

目的探讨葡萄糖转运蛋白1型(Glucose transporter—1，GLUT1)在脑肿瘤中的表达及其意义。方法收集55例颅脑手术标本，包括46例脑肿瘤患者和9例正常脑组织，采用单克隆抗体的GLUT1免疫组织化学SP法进行染色。结果7例低度恶性胶质瘤中的3例和全部13例高度恶性胶质瘤不同程度的表达GLUT1；10例良性脑膜瘤中的3例和全部5例恶性脑膜瘤为阳性表达；6例垂体瘤中的3例和5例表皮样囊肿也异常表达：所有正常脑组织约未见表达。结论四种脑肿瘤均可不同程度表达GLUT1，尤其在恶性肿瘤为明显，其与脑肿瘤的恶性程度相关。     

人脑肿瘤已糖激酶的活性测定

测定10例良性胶质瘤、18例恶性胶质瘤、12例良性脑膜瘤和1例恶性脑膜瘤的溶解性和总体己糖激酶(Hexokinase,HK)活性,以18例正常脑组织作对照.结果表明:良性与恶性脑瘤的HK活性明显低于正常脑组织(P<0.01).良性脑瘤的总体HK活性明显低于恶性脑瘤(P<0.05),脑膜瘤的HK活性明显低于良性及恶性胶质瘤(P<0.05,P相似文献   

非典型脑膜瘤

正2016年世界卫生组织(WHO)中枢神经系统肿瘤分类将非典型脑膜瘤定义为良性和恶性脑膜瘤中间类型,属WHOⅡ级。组织学形态观察,胞核分裂活性增强,肿瘤侵犯周围脑组织,或具备肿瘤细胞密度高、小细胞伴核质比高、核仁明显、不规则或片状生长、局灶自发性坏死中的3个及以上特点,其中,脑组织浸润为2016年最新定义,以肿瘤细胞呈不规则舌状浸润周围脑组织为特点(图1),肿瘤组织与脑组织之间无软脑膜,脑组织浸润常引起星形胶质细胞增生。免疫组织化学染色可见肿瘤组     

IL-13RoL2在脑胶质瘤中的表达及其临床诊断意义

目的探讨自细胞介素13受体α2（IL-13Rα2）在脑胶质瘤中的表达及其与肿瘤恶性程度的关系。方法免疫组化染色检测59例脑胶质瘤、3例脑膜瘤、3例癫痫病灶组织、5例正常脑组织及U251细胞中IL-13Rα2蛋白的表达,分析其与肿瘤恶性程度的关系。结果正常脑组织、脑膜瘤及癫痫病灶中均无IL-13Rα2的表达;而59例胶质瘤组织中33例IL-13Rα2表达阳性,阳性率55．9％;胶质瘤组织IL-13Rα2阳性表达率显著高于非胶质瘤组织（P〈0．05）。高级别胶质瘤（Ⅲ、Ⅳ级）IL-13Rα2阳性表达率为78．1％（25／32）,低级别（Ⅰ、Ⅱ级）胶质瘤为29．6％（8／27）,两者相较差异显著（P〈0．05）。U251细胞IL-13Rα2表达阳性。结论本结果提示胶质瘤中IL-13Rα2蛋白表达与其恶性程度有关。     

脑膜瘤缺氧诱导因子-lα表达与瘤周血管生成及病理级别的关系及意义

目的探讨脑膜瘤缺氧诱导因子-lα(HIF-lα)表达与瘤中血管生成及病理级别之间的关系,探讨HIF-lα在脑膜瘤发生和发展中的作用,为治疗脑膜瘤提供理论依据。方法选取脑膜瘤标本45例,正常脑组织标本10例(均为重型颅脑损伤急症行内减压术)作为对照。采用免疫组化ElivisionTMplus两步法,检测缺氧诱导因子-lα在脑膜瘤标本和正常脑组织标本中的表达,以人原始造血细胞(CD34)标记肿瘤微血管,用微血管密度(MVD)作为血管生成指标测定肿瘤血管生成,比较缺氧诱导因子-lα表达与血管生长及病理级别关系。结果45例脑膜瘤组织切片中有23例表达HIF-1α,染色阳性率为51.1%,HIF-1α在正常脑组织及脑膜瘤Ⅰ级、Ⅱ级、Ⅲ级中的阳性表达率分别为0.00%、32.00%、66.67%和87.50%。在各个级别的脑膜瘤及正常脑组织中,HIF-1α的阳性表达率间差异有显著性(P<0.05)。正常脑组织组、I、II和III级脑膜瘤组中的MVD值分别为10.34±2.78,17.25±2.69,20.43±5.20和30.79±5.64,MVD值随着脑膜瘤级别的升高逐渐增加。HIF-lα表达程度与MVD呈正相关。结论缺氧诱导因子-lα表达强度与脑膜瘤血管生成可能有关,为治疗脑膜瘤提供了新的思路。     

人脑胶质瘤中血管内皮生长因子表达与胶质瘤微血管数及肿瘤增殖活性关系的研究

目的　研究血管内皮生长因子 (VEGF)表达与胶质瘤微血管数、肿瘤增殖活性及其与胶质瘤恶性程度的相关性。方法　应用免疫组化方法检测 5 0例胶质瘤、8例正常脑组织中VEGF、血小板内皮细胞粘附分子 (CD3 1)和增殖细胞核抗原 (PCNA)的表达 ,测定其阳性细胞数和阳性血管数。结果　胶质瘤中均有VEGF、CD3 1及PCNA表达。其表达水平与肿瘤恶性程度呈显著正相关 ,其r分别为 0 .745 ,0 .765及 0 .685。结论　VEGF、CD3 1和PCNA在胶质瘤中的高表达是胶质瘤恶性表型之一 ,可作为病理诊断的补充。     

人脑胶质瘤转化生长因子α基因表达的研究

目的：研究转化生长因子α（ＴＧＦα）在胶质瘤中的表达及与肿瘤恶性程度的关系。方法：用Ｎｏｒｔｈｅｒｎ杂交和免疫组化法检测５０例胶质瘤，４例胶质瘤细胞系和８例正常脑组织的ＴＧＦαｍＲＮＡ表达与免疫反应性ＴＧＦα（ｉｈＴＧＦα）的表达水平。结果：正常脑组织表达极低水平的ＴＧＦαｍＲＮＡ，而胶质瘤组织与细胞系表达高水平的ＴＧＦαｍＲＮＡ。正常脑组织中未见ｉｈＴＧＦα阳性染色细胞，而胶质瘤组织与细胸系ｉｈＴＧＦα呈过表达。随肿瘤恶性程度增高，ＴＧＦαｍＲＮＡ和ｉｈＴＧＦα表达量亦增高。结论：胶质瘤中存在ＴＧＦαｍＲＮＡ和ｉｈＴＧＦα异常高表达，其表达水平可作为判断胶质瘤恶性度的一项客观指标。     

肌腱生长蛋白在人脑胶质瘤中的表达与增殖性细胞核抗原的相关性研究

目的 探讨肌腱生长蛋白（Tenascin,TN)和增殖性细胞核抗原（Proliferation cell nuclear antigen,PCNA)在不同级别人脑胶质瘤中的表达情况。分析它与胶质瘤的侵袭相关性。方法 采用免疫组织化学方法，检测54例不同级别人脑胶质瘤组织中Tenascin和PCNA的表达。结果 Tenascin表达于胶质瘤新生血管，瘤细胞周围基质以及胶质瘤细胞；PCNA表达于细胞核，随着胶质瘤病理级别的升高，肿瘤新生血管和瘤细胞周围基质中的Tenascin表达增强，瘤细胞中PCNA表达亦增强，Tenascin与PCNA的表达呈正相关。结论 Tenascin在胶质瘤血管形成，瘤细胞增殖转移过程中发挥了重要作用，Tenascin和PCNA可作为评价胶质瘤生物学行为的有用指标。     

Expression of tenascin in human gliomas: its relation to histological malignancy,tumor dedifferentiation and angiogenesis

Summary The immunohistochemical distribution of tenascin (TN), fibronectin (FN), and laminin (LN) was investigated in 56 human gliomas (8 astrocytomas, 15 anaplastic astrocytomas, and 33 glioblastomas) with regards to the histological degree of malignancy and the degree of tumor cell differentiation evaluated by the staining of glial fibrillary acidic protein (GFAP). In 8 anaplastic astrocytomas and 28 glioblastomas, TN was predominantly immunolocalized in the basement membrane zone of the proliferating tumor vessels; sections of all astrocytomas were negative for TN staining. FN was localized in the basement membrane zone of the vessels in all astrocytomas, 12 anaplastic astrocytomas, and 22 glioblastomas. In 7 anaplastic astrocytomas and 19 glioblastomas, both TN and FN were expressed to various degrees in the tumor vessels. However, most of the TN-positive vessels did not express FN, and most of the FN-positive vessels were negative for TN staining. Furthermore, in 6 anaplastic astrocytomas and 12 glioblastomas, either TN of FN, but not both, were expressed in any area on serial sections. Most of the tumor cells around TN-positive, FN-negative tumor vessels did not express GFAP. On the other hand, GFAP was present in most tumor cells around TN-negative, FN-positive vessels. LN was detected in all vascular and pial-glial basement membrane zone of the tissues examined. These findings indicate that the degree of histological malignancy and the degree of cell dedifferentiation of human gliomas correlate well with the expression of TN, but are inversely correlated with the expression of FN. We postulate that the expression of TN, but not of FN, plays a role in the promotion of angiogenesis in malignant gliomas.     

Elastase expression by infiltrating neutrophils in gliomas

Polymorphonuclear neutrophil elastase is a neutral protease released by activated polymorphonuclear neutrophils and plays a crucial role in maintaining host defense. However, under certain conditions, this enzyme damages normal tissue and facilitates infiltration of tumor cells. In this study, surgical specimens were obtained from the tumor core and infiltrating margin of the glioma of 12 patients with astrocytoma of varying degrees of malignancy, and the specimens were tested for the presence of elastase by immunohistochemical analysis. Polymorphonuclear neutrophil elastase was not present in the tumor core of any of the 12 cases. Elastase was expressed in areas of tumor infiltration of the brain in all four glioblastoma cases, three of the four anaplastic astrocytoma cases, and none of the four low-grade astrocytoma cases. There was a higher percentage of elastase-positive polymorphonuclear neutrophils in the infiltrating margin of tumors with greater degree of malignancy. Polymorphonuclear neutrophils are recruited to malignant gliomas, and the elastase released by these cells aids in the infiltration of gliomas.     

Glomeruloid blood vessels in ethylnitrosourea-induced rat gliomas

Summary Glomeruloid blood vessels (GBVs), a characteristic histological feature of most human malignant gliomas, were recognized with high incidence in autochthonous rat gliomas induced by transplacental administration of ethylnitrosourea. To evaluate some of the biological properties of these GBVs, we carried out a study using histological methods and immunohistochemical staining for glial fibrillary acidic protein, factor VIII-related antigen (VIII Ag) and bromodeoxyuridine (BrdUrd). Of 22 animals with large, massively growing gliomas in the CNS, GBVs including conglomerate aggregations of small blood vessels with endothelial hyperplasia and strong VIII Ag expression were observed in 13 large gliomas histologically consisting of primitive neuroepithelial neoplasms (PNN; so called ependymoma) and mixed-type gliomas in combination with astrocytoma and PNN or anaplastic astrocytoma. The anaplastic gliomas in our series were devoid of GBVs. These findings indicate that GBV formation takes place in a histological variety of experimental gliomas. Furthermore, the GBVs were frequently associated with the vasculo-mesenchymal stroma in the parent gliomas, suggesting an intimate relationship with the morphogenesis of GBVs. In addition, it was shown that the GBVs had a higher BrdUrd-labelling index than that of other blood vessels in gliomas and also that of neoplastic cells in most parent gliomas, except for anaplastic gliomas. Based on these results, the possible mechanism of GBV morphogenesis is discussed with regard to the roles of macromolecules in the induction and regulation of GBVs.Supported in part by grants nos. 63570675 and 01480350 from the Ministry of Education, Science and Culture, Japan, and by a grant from the Ministry of Health and Welfare, Japan     

Increase in levels of plasminogen activator and type-1 plasminogen activator inhibitor in human breast cancer: possible roles in tumor progression and metastasis

We measured antigen levels of two kinds of plasminogen activators, tissue type plasminogen activator (t-PA) and urokinase type plasminogen activator (UK), as well as their primary inhibitor, type-1 plasminogen activator inhibitor (PAI-1) in the tissue extracts of benign and malignant breast tumors. Tumor tissues of 36 fibroadenomas and 39 breast cancers were examined. t-PA levels were not different in both groups. Malignant tumors contained the significantly higher levels of UK than benign tumors (p less than 0.001). Furthermore in breast cancer tissues, UK antigen levels of tumors with axillary lymph node involvements were significantly higher than those of tumors without lymph node involvements (p less than 0.05). PAI-1 antigen levels of breast cancer tissues were dramatically higher than those of fibroadenoma (p less than 0.001). PAI-1 levels of node positive carcinomas showed also values significantly higher than node negative ones (p less than 0.01). When we divided cancer tissues into three groups as node negative tumors, tumors with positive axillary nodes fewer than four and tumors with four or more positive nodes, PAI-1 levels increased corresponding to the progression of lymph node involvements (p less than 0.05). Immunohistochemical studies, using mouse monoclonal antibodies to human UK and PAI-1, showed that those immunoreactivities were diffusely distributed in the cytoplasm of human breast cancer cells. Their staining patterns were very similar to each other.     

Progesterone regulation of plasminogen activator inhibitor 1 (PAI-1) antigen and mRNA levels in human endometrial stromal cells.

Plasminogen activator activity decreases in the endometrium in the secretory phase of the menstrual cycle. This is partly due to decreased release of urokinase plasminogen activator in response to progesterone. Plasminogen activator inhibitor type 1 (PAI-1) is an efficient inhibitor of both tissue-type and urokinase-type plasminogen activators, and may therefore be instrumental for the control of plasminogen activation. In this study we examined the effects of steroid hormones on PAI-1 release and PAI-1 mRNA levels in primary cultures of human endometrial stromal cells. In these cells the secretion of PAI-1 was increased by progesterone in a dose and time dependent way, but was not affected by estradiol. The progesterone induction of PAI-1 secretion was preceded by a 7-8 fold increase of the steady state level of PAI-1 mRNA in the cells, suggesting that progesterone activates PAI-1 gene expression. Cultured endometrial glandular epithelial cells were found to release only insignificant amounts of PAI-1 with or without hormone treatment. The effect of progesterone on endometrial stromal cells was mimicked by DH-testosterone. However, while the response to progesterone was completely blocked by ZK112993, a potent antagonist of the progesterone receptor, the response to DH-testosterone was partially blocked by ZK112993, and partially by OH-flutamide, a potent antagonist of the androgen receptor. This suggests that a secretory response on PAI-1 expression is mediated via androgen receptors in endometrial tissue.     

Deficient release of plasminogen activator inhibitor-1 from astrocytes triggers apoptosis in neuronal cells.

Plasminogen activator inhibitor-1 (PAI-1) plays an important role in the processes of peripheral tissue remodeling and fibrinolysis through the regulation of plasminogen activation. We found that cultured human astrocytes efficiently released PAI-1, and that both mRNA expression and protein release of PAI-1 were suppressed by pretreatment of the cells with daunorubicin. To examine the role of PAI-1 in the nervous system, neuronally differentiated PC-12 cells (PC-12 neurons) were maintained in a PAI-1-deficient culture medium derived from daunorubicin-pretreated astrocytes. The deficiency of PAI-1 in the medium caused a significant reduction in Bcl-2 and Bcl-XL mRNAs and an increase in Bcl-XS and Bax mRNAs in PC-12 neurons at 3 h. The changes in balance between mRNA expressions of the anti- and pro-apoptotic Bcl-2 family proteins caused caspase-3 activation following the release of cytochrome c from mitochondria. Apoptotic morphological change and DNA fragmentation were also observed in the neuronal cells at 24 h. Addition of exogenous PAI-1 protein to the inhibitor-deficient medium blocked the apoptotic changes in PC-12 neurons. However, addition of PAI-1 antibodies to control medium caused similar apoptotic changes in PC-12 neurons. During the apoptotic processes, plasminogen activator (PA) activity in the PAI-1-deficient medium was as low as the control level. The present data suggest that PAI-1 has physiological functions other than its role as PA inhibitor for the survival of neurons.     

Impaired capacity for upregulation of MHC class II in tumor-associated microglia

Immunotherapy for malignant gliomas is being studied as a possible adjunctive therapy for this highly fatal disease. Thus far, inadequate understanding of brain tumor immunology has hindered the design of such therapies. For instance, the role of microglia and macrophages, which comprise a significant proportion of tumor-infiltrating inflammatory cells, in the regulation of the local anti-tumor immune response is poorly understood. To study the response of microglia and macrophages to known activators in brain tumors, we injected CpG oligodeoxynucleotide (ODN), interferon-gamma (IFN-gamma), and IFN-gamma/LPS into normal and intracranial RG2 glioma-bearing rodents. Microglia/macrophage infiltration and their surface expression of MHC class II B7.1 and B7.2 was examined by flow cytometry. Each agent evaluated yielded a distinct microglia/macrophage response: CpG ODN was the most potent inducer of microglia/macrophage infiltration and B7.1 expression, while IFN-gamma resulted in the highest MHC-II expression in both normal and tumors. Regardless of the agent injected, however, MHC-II induction was significantly muted in tumor microglia/macrophage as compared with normal brain. These data suggest that microglia/macrophage responsiveness to activators can vary in brain tumors when compared with normal brain. Understanding the mechanism of these differences may be critical in the development of novel immunotherapies for malignant glioma.     

Elastase expression by infiltrating neutrophils in gliomas

Abstract

Polymorphonuclear neutrophil elastase is a neutral protease released by activated polymorphonuclear neutrophils and plays a crucial role in maintaining host defense. However, under certain conditions, this enzyme damages normal tissue and facilitates infiltration of tumor cells. In this study, surgical specimens were obtained from the tumor core and infiltrating margin of the glioma of 72 patients with astrocytoma of varying degrees of malignancy, and the specimens were tested for the presence of elastase by immunohistochemical analysis. Polymorphonuclear neutrophil elastase was not present in the tumor core of any of the 12 cases. Elastase was expressed in areas of tumor infiltration of the brain in all four glioblastoma cases, three of the four anaplastic astrocytoma cases, and none of the four low-grade astrocytoma cases. There was a higher percentage of elastase-positive polymorphonuclear neutrophils in the infiltrating margin of tumors with greater degree of malignancy. Polymorphonuclear neutrophils are recruited to malignant gliomas, and the elastase released by these cells aids in the infiltration of gliomas [Neurol Res 2000; 22: 465-468]