脑胶质瘤EGFR,p53蛋白和PCNA的表达与生物学行为

本文联合检测了５８例脑胶质瘤ＥＧＦＲ，ｐ５３蛋白和ＰＣＮＡ的表达，结果表明，三者的表达率均随胶质瘤的级别升高而增高增强，高级别组与低级别组相比较，有显著差异，复发与未复方组相比较，有显著差异，提示肿瘤的生物学行为与表达的高低有关，肿瘤的生物学行为越差，表达率就超高，染色强，预后越差。     

胶质瘤egr-1基因表达水平与其细胞增殖和凋亡关系的研究

目的：探讨胶质瘤细胞egr-1基因表达水平与肿瘤恶性程度、细胞增殖活性及凋亡程度的关系。方法：用原位杂交、原位细胞凋亡检测和免疫组化染色方法观察了73例不同级别的胶质瘤。结果：73例胶质瘤egr-1mRNA和EGR-1蛋白阳性表达率均为100％．这两种阳性肿瘤细胞密度均随肿瘤恶性程度升高而相应增加，不同级别组间比较差异均有显著性（P〈0．01）。73例胶质瘤增殖细胞核抗原（PCNA）阳性肿瘤细胞和凋亡肿瘤细胞检出率均为100％。随肿瘤恶性程度升高，PCNA阳性肿瘤细胞密度增加而凋亡肿瘤细胞密度减少，不同级别组间比较差异均有显著性（心0．05～0．01）。经直线相关分析证实，egr-1mRNA、EGR-1蛋白和PCNA阳性肿瘤细胞密度彼此间均呈显著性正相关（r=0．685～0．999，P〈0．01），前三种阳性肿瘤细胞密度均与凋亡肿瘤细胞密度呈显著性负相关（r=-0．758—0．775，P〈0．01）。结论：egr-1基因表达水平对评价胶质瘤生物学行为有重要参考价值。胶质瘤细胞egr-1基因表达异常增加可能是促进肿瘤细胞增殖和抑制其凋亡的重要因素，并在胶质瘤发生及恶性进展过程中均起重要作用。     

p15和WIF－1在人脑胶质瘤中的表达及其临床意义

目的：探讨p15和WIF－1在人脑神经胶质瘤中的表达,及其与临床病理特征和生物学行为的关系。方法：应用免疫组化SP法检测53例神经胶质瘤和20例正常脑组织中p15和WIF－1蛋白的表达,结合病例随访资料分析其临床意义。结果：p15及WIF－1蛋白在神经胶质瘤中的阳性表达率明显低于正常脑组织,差异具有统计学意义（P＜0．05）;p15及WIF－1蛋白的阳性表达率与神经胶质瘤的病理学分级和瘤周脑水肿具有相关性（P＜0．05）,但与性别和年龄不相关（P＞0．05）。p15和WIF－1蛋白在神经胶质瘤中的阳性表达率复发组与无复发组有明显差异（P＜0．05）。p15与WIF－1蛋白的表达呈正相关（r＝－0．396,P＜0．01）。结论：p15、WIF－1的表达与人神经胶质瘤的发生及进展相关;p15、WIF －1低表达患者预后较差。联合检测p15、WIF－1蛋白的表达对神经胶质瘤的早期诊断、判断预后有参考价值。     

胶质瘤细胞端粒酶基因表达对其增殖及凋亡影响的研究

背景与目的：端粒酶逆转录酶（hTERT）和端粒酶相关蛋白1（hTP1）是人端粒酶的重要组成部分。已知胶质瘤细胞有端粒酶异常再活化，而hTERT和hTP1基因表达是否也相应增加，尚不清楚。本研究是为了探讨胶质瘤细胞hTERT和hTP1基因表达、增殖活性及凋亡程度变化的相互关系及其在胶质瘤发生、发展中的作用。方法：采用mRNA原位杂交、免疫组织化学染色及原位细胞凋亡检测（TUNEL）等方法观察了70例不同级别的人胶质瘤组织标本。结果：本组70例胶质瘤hTERT mRNA和蛋白的阳性表达率分别为88．6％和82．9％．WHO Ⅰ～Ⅱ级组、Ⅲ级组及Ⅳ级组间比较二者的阳性表达率均无显著性差异（P〉0．05），hTP1 mRNA和蛋白及增殖细胞核抗原（PCNA）蛋白的阳性表达率均为100％。以上五种阳性肿瘤细胞的密度彼此间均呈显著性正相关（r=0．589～0．882，P〈0．0005），并均随肿瘤级别升高而相应增加，不同级别组间比较差异均有显著性（P〈0．05～0．01）。凋亡肿瘤细胞的检出率也为100％，但凋亡肿瘤细胞密度随肿瘤级别升高而相应减少，不同级别组间比较差异均有显著性（P〈0．01），且与前五种阳性肿瘤细胞密度间均呈显著性负相关（r=-0．551 ～-0．775，P〈0．0005）。结论：以上指标对评价胶质瘤的生物学行为均有重要参考价值，胶质瘤细胞hTERT和hTP1基因异常表达是其端粒酶再活化的先头条件，后者通过抑制肿瘤细胞衰老凋亡和促进其增殖在胶质瘤的发生、发展过程中均起重要作用。     

增殖细胞核抗原与p53蛋白在胶质瘤中的表达意义

本文采用PCNA和p53蛋白对45例胶质瘤进行联合探讨，发现两种核内蛋白均随肿瘤病理级别的升高而标记阳性升高。Ⅰ级组，未复发组的阳性率与Ⅱ级组、Ⅲ～Ⅳ级组、复发组相比较有显著差异（p＜0.01）．这提示肿瘤分级与病人的预后有关。本实验还证实胶质瘤的PCNA和p53蛋白易在细胞浆内表达，说明胶质瘤核与胞浆间有一定物质交换，提示可作为阳性判断．     

MMP-9、CD133蛋白在复发人脑胶质瘤中的表达及意义

背景与目的：肿瘤干细胞在胶质瘤复发中的作用备受关注,然而其获得侵袭力的具体机制尚不清楚。本课题旨在研究基质金属蛋白酶一9（MMP．9）、肿瘤干细胞标志物CDl33蛋白在复发人脑胶质瘤中的表达及可能作用。方法：采用免疫组织化学方法检测24例脑胶质瘤患者原发、复发两次手术组织标本及15例正常脑组织标本中MMP．9、CDl33蛋白的表达情况,并行相关性分析。结果：正常脑组织标本中未检测到MMP．9、CD133蛋白表达。24例脑胶质瘤患者两次手术标本中．CDl33和MMP-9蛋白表达强度均随着胶质瘤病理级别的增加而增高（氏0．01）。胶质瘤原发、复发两次手术组织标本中CDl33和MMP．9蛋白表达强度差异有统计学意SL（P〈0．05）：复发胶质瘤病理级别有增高趋势（P〈0．01）。CDl33和MMP-9蛋白表达呈正相关（P〈0．01）。结论：MMP．9、CDl33蛋白的表达与脑胶质瘤的恶性程度密切相关．可能在脑胶质瘤的复发过程中起协同作用。     

Ki-67和微血管密度在膀胱移行细胞癌中的表达及其相互关系

目的：探讨Ki-67抗原和微血管密度（MVD）在膀胱移行细胞癌中的表达及临床意义，并评价两者的相互关系。方法：利用鼠抗人Ki-67抗原单克隆抗体和鼠抗人FⅧ因子相关抗原（FⅧRAg）单克隆抗体对65例膀胱移行细胞癌和8例正常膀胱黏膜进行免疫组化染色。结果：Ki-67与MVD在癌组织中表达均显著高于正常黏膜（P＜0．01）。肿瘤中Ki-67指数和MVD之间存在正相关性，两者的表达均与膀胱移行细胞癌的病理分级显著相关，Ⅰ级与Ⅱ级、Ⅲ级与Ⅳ级之间存在显著差异（P＜0．01）；浸润性肿瘤组高于表浅肿瘤组（P＜0．01）；术后2年随访复发组高于未复发组（P＜0．01）。MVD与肿瘤大小有关，肿瘤直径大于2cm组MVD高于肿瘤直径小于2cm组（P＜0．05）。结论：Ki-67的表达为膀胱移行细胞癌的恶性表型，且与膀胱肿瘤血管形成有关。Ki-67指数和MVD对评价膀胱移行细胞癌的生物学形为和预后判断具有重要意义。     

胶质瘤DNA倍体、SPF值及EGFR蛋白表达的研究

目的：检测胶质瘤DNA倍性、SPF及EGFR表达，探讨其与胶质瘤病理级别的关系及它们之间相关性。方法：46例手术切除的脑胶质瘤作为实验组；10例正常脑组织作为对照组。采用免疫组化SABC（Strept-Avidin-Bion Complex)法检测EGFR蛋白表达；采用流式细胞技术（Flow-Cytometry,FCM)检测DNA含量及SPF。结果：对照组均为二倍体且无EGFR表达；胶质瘤I级组、Ⅱ级组、Ⅲ－Ⅳ级组DNA异倍体率分别为13．3％、46．2％、77．8％，I级组与Ⅲ－Ⅳ级组、Ⅱ级组与Ⅲ－Ⅳ级组比较差异有显著性（P＜0．05）；EGFR阳性率分别为20．0％、69．2％和83．3％，I级组与Ⅱ级组、I级组与Ⅲ－Ⅳ级组比较差异有显著性（P＜0．05）；SPF值随胶质瘤级别增高而升高，各组比较差异有显著性（P＜0．05）；EGFR阳性胶质瘤DNA发生率及SPF值均明显高于EGFR阴性胶质瘤（P＜0．05）。结论：DNA异倍体的发生、SPF值、EGFR阳性率与胶质瘤恶性级别正相关。EGFR表达与DNA异倍体及SPF有相关性。     

ING4及SKP2在不同级别脑胶质瘤组织中的表达

[目的]研究抑癌基因ING4及癌基因SKP2在正常脑组织及不同级别胶质瘤组织中的表达．以及其表达水平与脑胶质瘤生物学特征的关系。[方法]采用免疫组织化学技术检测70例手术切除的不同级别胶质瘤组织人及正常脑组织中ING4及SKP2的表达情况。[结果]ING4主要表达于细胞核和／或浆，正常脑组织中ING4的表达明显高于低级别与高级别胶质瘤，差异显著（t=11．8及22．5，P〈0．05）；低级别胶质瘤组织中ING4的表达明显高于高级别胶质瘤，差异显著（t=-17．9，P〈0．05）。SKP2主要表达于细胞核，部分为胞浆，正常脑组织中SKP2的表达明显低于低级别与高级别胶质瘤，差异显著（t=13．7及29．5，P〈0．05）；低级别胶质瘤组织中SKP2的表达明显低于高级别胶质瘤，差异显著（t=16．5，P〈0．05）。[结论]ING4基因在胶质瘤组织中表达明显下调，且ING4表达下调与胶质瘤的分级有密切关系；SKP2基因在胶质瘤组织中表达显著上调，且SKP2表达上调与胶质瘤的分级有密切关系。     

着丝粒蛋白F在脑胶质瘤中的表达及其意义

目的：检测着丝粒蛋白F（CENP—F）在脑胶质瘤中的表达，探讨其临床意义。方法：采用免疫组化方法检测64例脑胶质瘤和10例正常脑组织石蜡包埋标本中CENP—F的表达。结果：CENP—F在64例脑胶质瘤中表达阳性率为76．6％．在10例正常脑组织中仅30．0％，差异有统计学意义（P〈0．01）。CENP—F的表达在不同性别、年龄、肿瘤部位、肿瘤直径、瘤周水肿及病理分类中差异无统计学意义（P〉0．05），在不同病理分级中差异有统计学意义（P〈0．05），高分级组较低分级组表达高。结论：CENP—F在脑胶质瘤中高表达，与肿瘤病理分级有关，对判断脑胶质瘤生物学特性有一定意义。     

MDM2 and p53 expression in gliomas: a multivariate survival analysis including proliferation markers and epidermal growth factor receptor.

p53 and the murine double minute 2 (MDM2) oncoprotein expression was evaluated in paraffin-embedded tissue from 61 patients with central nervous system gliomas (53 astrocytomas and eight oligodendrogliomas) and related to proliferation-associated markers [i.e. proliferating cell nuclear antigen (PCNA), Ki-67 and nuclear organizer regions (NORs)] and epidermal growth factor receptor (EGFR). We used the monoclonal antibodies PC-10, MIB-1, DO-1, 1B1O and EGFR 113 and the colloid silver nitrate (AgNOR) technique. MDM2 and p53 were co-expressed in 28% of cases. A p53-positive/MDM2-negative phenotype was observed in 15% and a p53-negative/MDM2-positive phenotype in 20% of cases. There was a positive correlation of p53 and MDM2 expression with grade and proliferation indices. Univariate analysis in the group of diffuse astrocytomas showed that older age, high histological grade, high PCNA labelling index (LI) and high AgNOR score were associated with reduced overall survival (P < 0.05). p53 LI, Ki-67 LI, AgNOR score, tumour location and grade influenced disease-free survival (P < 0.05), whereas the only parameters affecting post-relapse survival were histological grade and Ki-67 LI (P < 0.1). Multivariate analysis revealed that age, radiotherapy, PCNA LI and p53 LI were the independent predictors of overall survival. p53 LI, Ki-67 LI, MDM2 LI, EGFR LI, grade and type of therapy were independent predictors of disease-free survival, and grade was the only independent predictor of post-relapse survival. Our results indicate that p53 LI and MDM2 LI, EGFR expression as well as proliferation markers (PCNA and Ki-67) are useful indicators of overall and disease-free survival in diffuse astrocytoma patients.     

食管鳞癌中p53、PCNA和EGFR的表达与化疗疗效的关系

目的:探讨p53、PCNA和EGFR在食管鳞癌中的表达情况与化疗疗效的关系。方法:对66例食管鳞癌化疗前的活检标本用免疫组化技术分别检测p53、PCNA、EGFR的表达。结果:p53蛋白积聚阳性组化疗有效率(16.7%)明显低于p53蛋白积聚阴性组(70.8%)(P<0.01),PCNA过表达组化疗有效率(79.4%)高于PCNA弱表达组(31.3%)(P<0.01),EGFR过表达组化疗有效率(30.4%)低于弱表达组(69.8%)(P<0.01)。经Logistic回归分析,3个指标中,PCNA对化疗疗效的预测价值较大(P<0.01)。结论:食管鳞癌中p53、PCNA和EGFR的表达情况对化疗疗效均有一定的预测价值。其中PCNA价值较大。     

VEGF、p53、PCNA蛋白表达与垂体腺瘤侵袭性复发性关系的研究

目的探讨VEGF、p53、PCNA蛋白表达与垂体腺瘤发生及其生物学行为的关系.方法应用免疫组化染色方法检测VEGF、p53、PCNA蛋白在30例侵袭性垂体腺瘤及30例非侵袭性垂体腺瘤中的表达.结果垂体腺瘤VEGF、p53、PCNA 蛋白表达水平在侵袭组中明显高于非侵袭组,复发组明显高于非复发组.结论VEGF、p53、PCNA蛋白异常表达与垂体腺瘤的发生及其生物学行为改变有关,并可以作为肿瘤侵袭性的生物学指标.     

Correlation of chromosomal imbalances by comparative genomic hybridization and expression of EGFR, PTEN, p53, and MIB-1 in diffuse gliomas

The histological subclassification of gliomas is increasingly assisted by the underlying molecular genetics which has major importance in guiding clinical management of the disease. However, the assessment of several molecular events for improving clinical care remains a challenge. Herein, we report on comparative genomic hybridization (CGH) and immunohistochemical (IHC) assessment of EGFR, PTEN, p53, and MIB-1 expression in 13 oligodendrogliomas (10 WHO grade II, 3 WHO grade III), one oligoastrocytoma (WHO grade III) and 23 high-grade astrocytomas (3 WHO grade III, 20 glioblastoma multiforme). The most frequent imbalances in oligodendroglial tumors including the oligoastrocytic case were, in decreasing order of frequency, +7q, -1p, and -4q and in astrocytomas +7q, -10q, +7p, -9p, -10p, +20q, and +20p. Some individual imbalances were associated with increasing numbers of chromosomal changes, that were +7q in both oligodendrogliomas and astrocytomas, and -9p, -10q, +20p, and +20q in astrocytomas. The markers p53 and MIB-1 were significantly higher expressed in astrocytomas than in oligodendrogliomas and expression levels of p53 and EGFR were inversely associated within the astrocytic group. In addition, p53 overexpression correlated positively with +7q and negatively with -1p in the oligodendroglial group whereas EGFR overexpression correlated positively with -1p in the oligodendroglial and positively with +7p and -10p in the astrocytic group. Short overall survival was significantly associated with +7p and -10q in astrocytomas. Collectively, these results contribute to the increasing clinical relevance of assessing tumor biological markers in gliomas.     

Expression of biomarkers (p53, transforming growth factor alpha, epidermal growth factor receptor, c-erbB-2/neu and the proliferative cell nuclear antigen) in oropharyngeal squamous cell carcinomas

Using immunohistochemistry, expression of p53, transforming growth factor-alpha (TGF-alpha), epidermal growth factor receptor (EGFR), c-erbB-2/neu and proliferating cell nuclear antigen (PCNA) was examined in 26 fresh frozen tissue specimens of oropharyngeal squamous cell carcinomas (SCCs). p53 gene mutations were examined by polymerase chain reaction (PCR)/DNA sequencing methods in 22 carcinomas. The findings were examined for correlations with patients' clinicopathological parameters. Expressions of p53 and PCNA were also examined in 21 formalin-fixed corresponding tissues. Of the fresh frozen tissue specimens, 77% (20/26) showed expression and 68% (15/22) showed mutations (substitutions) of the p53, with significant clustering of the mutations in exons 5 (8/22; 36%), 7 (4/22; 18%) and 8 (5/22; 23%). No mutations were found in exon 6. There was a discordance between expression of p53 protein and mutations of the gene. Parallel to expression and mutations of the p53 found in most of the specimens, expression of TGF-alpha, EGFR, c-erbB-2/neu and PCNA was found in 88% (22/25), 92% (23/25), 58% (14/24) and 91% (21/23) of the specimens, respectively. For the formalin-fixed tissue specimens, 62% (13/21) and 90% (19/21) expressed p53 and PCNA, respectively. Examining for correlations with patients' clinicopathological parameters, expression of p53, TGF-alpha, EGFR and c-erB-2/neu seemed to negatively correlate with the increase of the tumour grade. The present work suggests that: (1) lack of negative growth regulation due to inactivation of the p53 gene together with activation of other proto-oncogenes are necessary genetic events in the carcinogenesis of oropharyngeal SCCs; (2) in oropharyngeal SCCs, p53 gene mutations were clustered in exons 5 (codons 130-186), 7 (codons 230-248) and 8 (codons 271-282) which perhaps suggests that tobacco carcinogens probably affect the mutational hot spots of the p53 gene at codons 157, 175, 186, 248, 273 and 282; and (3) fresh frozen and formalin-fixed tissue specimens give similar results when an immunohistochemical method is applied. The importance of p53, TGF-alpha, EGFR, c-erbB-2/neu and PCNA as biomarkers in oropharyngeal SCCs deserves particular attention because it might offer further understanding of the development of these carcinomas.     

脑胶质瘤组织p14^ARF、p53和p21^WAF1的表达及其意义

目的：探讨p14^ARF、p53和p21^WAF1蛋白在不同级别脑胶质瘤组织中的表达及其在胶质瘤发生、发展中的生物学意义。方法：应用免疫组化SP法检测Ⅱ级和Ⅳ级胶质瘤组织中p14^ARF、p53和p21^WAF1蛋白的表达,并分析其与胶质瘤组织学分级之间的关系。结果：Ⅱ级和Ⅳ级胶质瘤中,p53蛋白的阳性表达率分别为28．00％（7／25）及60．87％（14／23）（P=0．022）,其阳性表达率随胶质瘤恶性程度的增加而升高,Spearman等级相关分析显示,p53的表达与胶质瘤分级呈正相关（P〈0．05）;p21^WAF1的阳性表达率分别为76．00％（19／25）及39．13％（9／23）（P=0．010）,P14^ARF的阳性表达率分别为76．00％（19／25）及34．78％（8／23）（P=0．004）,二者阳性表达率均随胶质瘤恶性程度的增加而降低,Spearman等级相关分析显示,p21^WAF1、p14^ARF的表达与胶质瘤分级呈负相关（P〈0．05）。结论 胶质瘤组织中,p53呈不同程度的过表达,且随胶质瘤恶性程度的增加而表达水平升高;p21^WAF1、p14^ARF随胶质瘤恶性程度的增加表达水平降低。突变型p53蛋白的过表达以及p21^WAF1、p14^ARF蛋白的低表达,可促进胶质瘤的发生、发展。     

Biologic behavior of and p53 overexpression in multifocal renal cell carcinoma of clear cell type: an immunohistochemical study correlating grading, staging, and proliferation markers

BACKGROUND: In the treatment of small renal cell carcinoma (RCC), there is controversy between radical and nephron-sparing surgical treatment because of the risk of tumor multifocality. The biologic behavior of multifocal RCC compared with that of unifocal RCC is not well investigated, and the relevance of p53 and the proliferation markers MIB-1 and proliferating cell nuclear antigen (PCNA) to multifocal RCC is not yet established. METHODS: In this study, p53 protein overexpression was investigated immunohistochemically in 27 multifocal and 65 unifocal clear cell RCCs using a monoclonal antibody (DO-1). The nuclear expression of p53 was compared with the expression of PCNA and MIB-1 (Ki-67 antigen) and other prognostic factors, including grade and stage. RESULTS: Thirty-three RCCs (35.9%) had p53 positive nuclear staining. MIB-1 positivity was significantly higher in p53 positive tumors than in p53 negative tumors. PCNA positivity was not different in p53 positive tumors compared with p53 negative tumors. Proliferation marker expression was not associated with tumor focality. p53 overexpression was more often found in unifocal tumors than in multifocal tumors. Intracellular accumulation of the p53 protein was related to tumor grade but not to the T classification of tumor stage. In addition, lymph node involvement was significantly associated with p53 overexpression in tumors of the kidney. Focality did not influence progression free survival. CONCLUSIONS: This study demonstrated that there is no difference in the proliferative activity or biologic behavior of multifocal and unifocal tumors.     

鼻腔鼻窦鳞状细胞癌组织中PCNA和EGFR的表达

目的　探讨增殖细胞核抗原 (PCNA )和表皮生长因子受体 (EGFR )在鼻腔鼻窦鳞状细胞癌组织中的表达 ,并分析其与临床病理特征的关系。方法　采用免疫组化方法 ,对 45例鼻腔鼻窦鳞癌组织进行PCNA和EGFR检测 ,并用 15例正常鼻黏膜作对照。结果　PCNA和EGFR在各级鼻腔鼻窦鳞癌组织中均有表达 ,与正常鼻黏膜比较有显著性差异 ,均有随鳞癌分级的升高而表达增强的趋势。PCNA、EGFR在鼻腔鼻窦鳞癌中的表达有显著的相关性。结论　PCNA和EGFR的表达均能反映鼻腔鼻窦鳞癌的增殖活性 ,表达强度可反映鳞癌细胞增殖活性的高低     

肾盂癌p53蛋白、AgNOR和PCNA相关性及临床意义的研究

采用免疫组化和银染色技术对31例肾盂癌组织中p53蛋白，核仁组成区嗜银蛋白（AgNOR）和增殖细胞核抗原（PCNA）进行研究。发现肾盂癌p53表达阳性率，AgNOR颗粒计数和PCNA增殖指数均随肿瘤病理分级、临床分期的上升而增加，且与预后呈负相关。p53蛋白阳性表达组AgNOR颗粒计数和PCNA增殖指数显著高于p53蛋白阴性表达组。肾盂癌AgNOR计数与PCNA增殖指数间存在正相关性。提示p53蛋白表达，AgNOR计数和PCNA免疫染色在肾孟癌生物学行为及预后评估中具有重要意义。