人脑胶质瘤中表皮生长因子受体表达的研究

目的：探讨表皮生长因子受体（ＥＧＦＲ）基因表达与胶质瘤恶性程度的关系。方法：采用Ｎｏｒｔｈｅｒｎ印迹杂交及免疫组织化学技术从ｍＲＮＡ及蛋白水平检测了５０例人脑胶质瘤、４个体外恶性胶质瘤细胞系及８例正常脑组织的ＥＧＦＲ的基因表达。结果：免疫组化检测高恶度胶质瘤（ＷＨＯⅢ－Ⅳ级）ＥＧＦＲ表达６９％（２０／２９），而低恶度胶质瘤（ＷＨＯⅠ－Ⅱ级）表达率为３３％（７／２１），差异显著（Ｐ＜０．０１）；４例细胞系阳性表达１００％；８例正常脑组织无ＥＧＦＲ蛋白表达。ＷＨＯ分级与ＥＧＦＲ蛋白表达呈正相关（ｒ＝０．５５９７，Ｐ＜０．００１）。对５０例胶质瘤进行Ｎｏｒｔｈｅｒｎ印迹杂交，其结果与免疫组化检测结果一致，ＥＧＦＲｍＲＮＡ也随胶质瘤恶性程度的增加而表达增高。结论：ＥＧＦＲ可能在恶性胶质瘤的发生发展中起重要作用，ＥＧＦＲ过表达与胶质瘤分级相关性为从分子水平评估肿瘤的恶性程度及选择基因治疗的靶基因提供了参考。     

脑肿瘤中P53基因突变与突变型P53蛋白表达的研究

采用聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）法对４１例脑肿瘤进行Ｐ５３基因突变的检测，并与抗突变型Ｐ５３蛋白单抗的免疫组化染色结果相对照，结果显示脑肿瘤中Ｐ５３基因突变率为３４．１％（１４／４１），胶质瘤为２８％（１０／３６）。Ｐ５３基因突变与突变型Ｐ５３蛋白表达具有高度一致性，且二者均与脑肿瘤的分化程度有关。Ｐ５３基因突变及蛋白的表达不仅出现在高恶度胶质瘤及转移癌，而且可出现在低恶度胶质瘤中，表明Ｐ５３基因的突变在脑瘤的发生、发展过程中起重要作用。     

神经鞘瘤的schwannomin/merlin表达分析

目的 分析S／M蛋白在神经鞘瘤中的表达。方法 用免疫组织化学、蛋白印迹法观察36例神经鞘瘤中S／M的表达。结果 所有神经鞘瘤无Ⅱ型多发神经纤维瘤病（NF2）（A－19）免疫活性。28例（78％）S／M表达量下降（与正常比小于70％），其中听神经瘤占81％（13／16）、非听神经瘤占75％（15／20）。13例（36％）（听神经瘤3例、非听神经瘤10例）中等度下降（与正常比下降35％－69％）；15例（42％）（听神经瘤10例、其它5例）表达重度下降（与正常比小于35％）。结论 神经鞘瘤中S／M表达下降。     

脑和脊髓肿瘤患者脑脊液中唾液酸含量测定的临床意义（摘要）

脑和脊髓肿瘤患者脑脊液中唾液酸含量测定的临床意义（摘要）文芳，宋业胜，李承晏一、对象与方法１．对象：（１）病人组：脊髓肿瘤１７例（包括神经鞘瘤７例，胶质瘤１０例）。脑肿瘤１２例（包括脑膜瘤４例，胶质瘤８例）。均经病理切片确诊，中枢神经系统白血病９例经...     

小儿脑胶质瘤p16基因的表达

目的：检测２０例小儿脑软质瘤中Ｐ１６基因及其蛋白的存在情况。方法用聚合酶链式反应－单链构象多态性分析和免疫组织化学技术检测了２０例１－１４岁小儿脑胶质瘤。成果显示Ｐ１６基因和Ｐ１６蛋白的丢失率分别为５０％（１０／２０％）及（５／２０）。结论本结果提示Ｐ１６基因和蛋白缺失可能与部分小儿脑有质瘤发生,发展有关。     

人脑胶质瘤WAF1／CIP1表达的研究

目的检测抑癌基因ＷＡＦ１／ＣＩＰ１在人脑胶质瘤中的表达。方法取４６例胶质瘤组织及１２例脑外伤组织，提取其ｍＲＮＡ，用ＷＡＦ１／ＣＩＰ１作为模板进行逆转录ＰＣＲ（ＲＴ－ＰＣＲ）。结果４６例胶质瘤中有１４例有ＷＡＦ１／ＣＩＰ１蛋白表达（１４／４６），表达率为３０．４％，１２例脑外伤标本全部表达（１２／１２），表达率为１００％，两组比较有显著差异（Ｐ＜０．０１）。结论ＷＡＦ１／ＣＩＰ１的缺失是脑胶质瘤发生及进展的原因之一。     

听神经瘤BCL-2蛋白及bcl-2/JH融合基因的研究

目的 评价石蜡包埋听神经瘤组织中ＢＣＬ－２蛋白表达及相关的ｂｃｌ－２（ｍｂｒ）／ＪＨ融合基因改变,以探讨ｂｃｌ－２癌基因在听神经瘤发病中的可能意义。方法 免疫组化检测石蜡包埋组织中ＢＣＬ－２蛋白的表达;提取石蜡包埋组织的ＤＮＡ,ＰＣＲ检测ｂｃｌ－２（ｍｂｒ）／ＪＨ融合基因。结果 本组４０例听神经瘤,ＢＣＬ－２蛋白表达阳性２７例（６７．５％）,ｂｃｌ－２（ｍｂｒ）／ＪＨ融合基因检出阳性１９例（４７．５％）．结论 听神经瘤中存在ＢＣＬ－２蛋白的高表达及ｔ（１４：１８）染色体易位,提示雪旺氏细胞凋亡抑制可能是听神经瘤发病的分子病理基础之一。     

胶质瘤多药耐药细胞系C6／adr的建立及其耐药性逆转的研究

目的 研究胶质瘤多药耐药（ＭＤＲ）的发生机理及逆转方法。方法 建立了Ｃ６／ａｄｒ ＭＤＲ细胞系,经ＲＴ－ＰＣＲ及免疫组化染色分别研究了ｍｄｒ－１基因及Ｐ－糖蛋白（Ｐ－ｇｐ）的表达;采用ＭＴＴ药敏试验及ＨＰＬＣＡ测定细胞内阿霉素浓度的方法研究了异搏定、红霉素、潘生丁、Ｐ－ｇｐ单克隆抗体、复方丹参对ＭＤＲ的逆转作用。结果 Ｃ６／ａｄｒ ＭＤＲ细胞系ｍｄｒ－１基因阳性,Ｐ－ｇｐ高度表达。异搏定（２～６μ     

人脑胶质瘤细胞多药耐药性的形成及其耐药特性的研究

目的探讨人脑胶质瘤细胞多药耐药性（ＭＤＲ）的形成规律及其耐药特性。方法采用长春新碱（ＶＣＲ）在体外对人脑胶质瘤ＢＴ３２５细胞持续诱导获得了表现为ＭＤＲ特征的人脑胶质瘤细胞模型．以ＭＴＴ法测定胶质瘤细胞增殖，透射电镜行超微结构研究，通过流式细胞仪，检测培养的胶质瘤细胞与３２例术后胶质瘤新鲜组织标本中Ｐ－糖蛋白的表达。结果耐药的胶质瘤细胞ＢＴ３２５／ＶＣＲ对长春新碱的耐受程度为其亲本细胞的２４倍，对阿霉素、威猛交叉耐药，对５－氟脲嘧啶敏感。透射电镜下见ＢＴ３２５／ＶＣＲ细胞常染色质明显，线粒体丰富，粗面内质网增多。研究表明ＢＴ３２５／ＶＣＲ表现为Ｐ－糖蛋白介导的典型ＭＤＲ。胶质瘤新鲜组织标本中，Ｐ－糖蛋白表达阳性率为３７５％（１２／３２），同肿瘤的恶性程度呈正相关（Ｐ＜０．０１）。结论长春新碱能够诱导人脑胶质瘤细胞产生Ｐ－糖蛋白介导的典型ＭＤＲ。胶质瘤中多药耐药基因产物Ｐ－糖蛋白表达的阳性率同肿瘤的恶性程度呈正相关。应用流式细胞仅能早期发现耐药细胞，具有临床推广价值。     

鼻腔神经组织肿瘤临床病理与预后

３１例鼻腔神经组织肿瘤临床与病理分析表明：该类肿瘤是鼻腔内比较少见的肿瘤，可见于各年龄组。病理分类：嗅神经母细胞瘤１８例，黑色素瘤１例，脑膜瘤６例，胶质瘤４例，神经鞘瘤２例。良性肿瘤需手术切除，预后良好，恶性肿瘤不需要手术彻底且术后加用放射治疗。对１３例恶性肿瘤进行随访，其１年生存率（１２／１３）为９２．３％，３年生存率（７／１３）为３２．８％，５年生存率（３／１３）为２３．０８％。结合临床表现分     

Low frequency of replication errors in primary nervous system tumours

OBJECTIVES: Automated DNA technology was used to analyze the incidence of microsatellite instability (MIN) among the most frequent types of adult primary CNS tumours and to determine its relation with clinicopathological characteristics. METHODS: Fifty six gliomas, 32 meningiomas and 11 schwannomas were screened for size changes at eight microsatellite loci using fluorescent polymerase chain reaction (PCR) followed by fragment analysis in an automated sequencer. A tumour was considered as MIN+ when a different electrophoretic pattern between constitutional and tumour DNA was evidenced in one or more microsatellite markers and as replication error positive (RER+) when at least 25% of the markers analyzed (2/8) showed instability. The MIN phenotype was correlated with relevant clinical and pathological parameters. RESULTS: Globally, instability was found in 19/767 analyses (2.47%), with a higher rate among tetranuceotide than dinucleotide repeats (chi(2) test, p=0.018). Ten gliomas (17.9%), two meningiomas (6.3%), and two schwannomas (18.2%) were MIN+, whereas one glioma (1.8%), two meningiomas (6.3%), and one schwannoma (9.1%) were classified as RER+. A possible association between microsatellite instability and a shorter duration of clinical course was found in meningiomas. The MIN+ phenotype was more frequent in spinal than intracranial schwannomas (Fisher's exact test, p=0.018). No other significant association with clinical or histological features was detected. CONCLUSIONS: Although microsatellite instability can be demonstrated at a low rate in some primary CNS tumours, a true replication error phenotype (revealed by widespread microsatellite instability at numerous loci) is uncommon and unlikely to play an important part in the pathogenesis of these neoplasms. This form of instability was more frequent in tetranucleotide than in dinucleotide repeats. To our knowledge, this is the first report of MIN in schwannomas, where it was associated with the spinal localisation of the tumour.     

Expression of p67 (Munc-18) in adult human brain and neuroectodermal tumors of human central nervous system

p67 (Munc-18), is a neuron-specific protein of 67 kDa, known for its ability to bind with syntaxin and also to copurify with neuronal cdc2-like kinase. Earlier, in situ hybridization and immunocytochemical analysis of rat trigeminal ganglion and hippocampal cells demonstrated the specific localization of p67 in nerve cells and its rich distribution in axons. In the present study, we have looked for p67 expression in normal human brain and various neuroectodermal tumors. Immunohistochemical and Western immunoblot analysis of normal human brain tissue using antibodies against the N- and C-termini of p67 demonstrated the specific localization of this protein in postmitotic neurons but not in glia. Among neuroectodermal tumors, expression of p67 was observed in 100% of the tumors of neuronal origin studied, especially in the mature neuronal cell population of these tumors. Western immunoblot analysis of non-neuronal neuroectodermal tumors failed to reveal the expression of this protein in majority of cases. However, in gliomas and meningiomas, mild cytoplasmic immunohistochemical staining of neoplastic cells was noted in 64.7% and 25% of cases, respectively. Observed mild immunohistochemical staining of these tumors could be due to immunoreactivity to low molecular weight degraded products of p67, as seen on Western blot. The findings suggest that p67, by virtue of its ability to be expressed in postmitotic neurons of adult human brain and in tumors of neuronal origin, may serve as a molecular tool to understand the growth and differentiation of the nervous system in general. Received: 8 September 1998 / Revised: 23 December 1998, 1 March 1999 / Accepted: 12 April 1999     

人脑肿瘤染色体位点特异性杂合性丢失的研究

目的通过检测脑肿瘤杂合性丢失(LOH)的集中区域协助有关肿瘤抑制基因(TSG)的查找。方法选取9个微卫星多态标记,以PCR扩增-变性测序凝胶电泳检测67对瘤标本和相应正常组织DNA。结果胶质瘤在13q、19q和22q等位点存在较高频率(>20％)的LOH,脑膜瘤在22q和1p出现高频率(>30％)的LOH,神经鞘瘤仅在22q位点出现40％的LOH。结论脑肿瘤高频率的染色体位点特异性LOH与TSGs的失活密切相关。     

Expression of neurofibromatosis 2 protein in human brain tumors: an immunohistochemical study

The neurofibromatosis 2 (NF2) gene-encoded protein, named merlin, may function as a molecular linkage connecting cytoskeleton and plasma membrane. Merlin is thought to play a crucial role as a tumor suppressor not only in hereditary NF2-related tumors, but also in sporadic tumors such as schwannomas, meningiomas and gliomas. Using a merlin-expression vector system, we raised specific antiserum against merlin. We observed the intracellular distribution of merlin in cultured glioma cells, and further investigated merlin expression in 116 human brain tumors. Immunofluorescence microscopy revealed that merlin was localized beneath the cell membrane and concentrated at cell-to-cell adhesion sites, where actin filaments are densely associated with plasma membrane. By immunohistochemistry, none of the schwannomas from either NF2 patients or sporadic cases showed any immunoreactivity, while normal Schwann cells of cranial nerves were immunopositive. In meningiomas, merlin expression was frequently seen in the meningothelial subtype (8/10, 80%), but no expression could be detected in either the fibrous or the transitional variant. Most normal astrocytes were negative; however, reactive astrocytes often expressed merlin. Glioblastomas and anaplastic astrocytomas were found to be strongly positive, and focal positive staining was observed in fibrillary and pilocytic astrocytomas. Thus, the loss of merlin appears to be integral to schwannoma formation and the differential pathogenesis of meningioma subtypes. However, merlin alterations do not appear to play a critical role in either the tumorigenesis or malignant transformation of neoplastic astrocytes. Received: 8 July 1996 / Revised, accepted: 23 September 1996     

Immunostaining for proliferating cell nuclear antigen: its role in determination of proliferation in routinely processed human brain tumor specimens

Alcohol- and formalin-fixed, paraffin-embedded samples of 71 brain tumors (35 gliomas, 22 metastatic carcinomas, 8 meningiomas and 6 other tumors) were investigated by immunocytochemistry with three different monoclonal antibodies against proliferating cell nuclear antigen (PCNA)/cyclin (19A2; 19F4; PC10). PC10 was found to work best; it is applicable to both alcohol- and formalin-fixed tumor samples. PCNA labeling indices (LIs) were compared in the same tumors with LIs obtained by Ki-67 immunostaining of frozen sections and by in vitro incubation with bromodeoxyuridine (BrdUrd); in the latter preparations, BrdUrd LIs could be compared with PCNA LIs in the very same areas of serial sections. In gliomas, PCNA LIs were 0.7–80.2% (mean 31.7%), in metastases 0–76.0% (mean 47.8%), and in meningiomas 0–53.0% (mean 19.3%). In general, PCNA LIs were highly significantly correlated with Ki-67 LIs (P=0.0002) and BrdUrd LIs (P=0.0001). However, when tumor subgroups are considered, only gliomas show a significant correlation with Ki-67 and BrdUrd LIs. Despite this statistical correlation, PCNA expression was out of proportion to proliferation indices as determined by both other methods in almost one third of all brain tumors. Immunocytochemistry for PCNA produces a broad spectrum of staining intensity of labeled nuclei, whose number is dependent upon the sensitivity of the immunocytochemical technique used. Thus, inter-oberserver and inter-laboratory variabilities in PCNA LI determination may occur. Overlapping of PCNA LIs between tumor subgroups of varying malignancy further limits the informational value for the individual case. In some classic meningiomas, high PCNA scores do not reflect the proliferative activity of the tumor, as Ki-67 and BrdUrd LIs are very low in these cases. We conclude that PCNA immunolabeling is of limited value in the individual tumor, mainly due to overexpression in many tumors, and at present cannot be recommended to replace Ki-67 and/or BrdUrd labeling methods for routine determination of proliferative activity in human tumor specimens.Supported in part by a grant from the Oncology Commission, Medical Faculty, University of Vienna     

Estrogen receptors in brain tumors

We examined the cytosolic estrogen receptor (ER) level in tumor tissue from 77 patients: 36 meningiomas, 20 gliomas (12 glioblastomas, 2 cerebellar astrocytomas, 2 ependymomas, and 4 medulloblastomas), 8 neurinomas, 7 pituitary adenomas (2 prolactin-producing adenomas, 1 growth hormone-producing adenoma, and 4 nonfunctioning adenomas), and 6 metastatic brain tumors (1 from breast cancer, 4 from lung cancers, and 1 from colon cancer). Nuclear ER levels were assayed in 11 meningiomas and 2 glioblastomas. ER was determined by the dextran-coated charcoal method and calculated by Scatchard analysis. Cytosolic ER was detected in 100% of the pituitary adenomas, 50% of the meningiomas, 50% of the metastatic brain tumors, 25% of the neurinomas, and 15% of the gliomas. In gliomas, only medulloblastomas had ER activity. Nuclear ER was found in three premenopausal women with meningioma. The dissociation constant of the ER complex was, in each case, less than 10(-9) M. These observations suggest that some brain tumors may be responsive to estrogen via the cellular ER.     

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   

100例脑肿瘤中p53基因突变与P53蛋白积聚的研究

目的:研究不同类型脑肿瘤中的p53基因突变与P53蛋白积聚及其相关性。方法:采用聚合酶链反应-单链构象多态性(PCR-SSCP)分析及免疫组化法检测100例脑肿瘤p53基因突变及蛋白表达。结果:p53基因突变率为11%(11/100),其中高恶度胶质瘤为37.5%(6/16),低恶度胶质瘤4.3%(1/23),脑膜瘤6.9%(2/29),转移瘤40.0%(2/5)。P53蛋白表达阳性率为22%(22/100),其中高恶度胶质瘤为62.5%(10/16),低恶度胶质瘤为26.1%(6/23),脑膜瘤10.3%(3/29),转移瘤60%(3/5);其他肿瘤均未发现p53基因突变或蛋白表达。P53蛋白表达阳性的22例中伴有p53基因突变者11例,多见于高恶度肿瘤。结论:p53基因失活在脑肿瘤恶性进展过程中起重要作用。p53基因突变与P53蛋白积聚相关,但并非唯一因素。     

Proliferation kinetics of human brain tumors by in situ double labeling with bromodeoxyuridine and iododeoxyuridine]

Nineteen patients with human brain tumors (9 gliomas, 6 metastatic brain tumors, 3 meningiomas, 1 neurinoma) received intravenous infusions of iododeoxyuridine (IUdR) and bromodeoxyuridine (BUdR) at different time sequences, to estimate the duration of S-phase (Ts) and the potential doubling time (Tp) of individual tumors. Excised tumor specimens were reacted with Br-3, a monoclonal antibody that identifies only BUdR, and with IU-4, a monoclonal antibody that recognizes both IUdR and BUdR, and then were stained immunohistochemically. The BUdR LIs varied from 0.9% to 26.0%, reflecting the malignancy of each tumor. Despite the difference in LIs, however, the Ts measured was fairly uniform. The Ts was 8.9 +/- 1.8 hrs (mean +/- SD) in malignant gliomas, 9.2 +/- 2.5 hrs (mean +/- SD) in metastatic brain tumors and 9.2 +/- 0. 3 hrs (mean +/- SD) in meningiomas, respectively. In contrast, the Tp varied from 1.3 to 12.4 days in malignant gliomas and from 1.2 to 4.4 days in metastatic brain tumors. Double logarithmic regression analysis showed a close correlation between the BUdR LI and Tp in human brain tumors with LI > 1%. Double labeling studies with BUdR and IUdR allow some of the proliferation characteristics to be determined from a single biopsy specimen and provide more useful information of each tumors than can be obtained by single-labeling studies with BUdR.