少突胶质瘤染色体1p/19q联合缺失与Ki-67表达的关系

目的观察少突胶质细胞瘤1号染色体短臂(1p)和19号染色体长臂(19q)联合杂合性缺失(LOH)与Ki-67蛋白表达的相关性,探索预测少突胶质瘤化疗敏感性的分子标记物。方法少突胶质瘤肿瘤组织标本31例作为实验组,少突胶质瘤瘤旁正常脑组织标本13例作为对照组,免疫组化方法检测两组Ki-67蛋白的表达,荧光原位杂交技术检测1p/19q的缺失情况。结果对照组有1例Ki-67表达呈现阳性(7.7%),实验组有14例Ki-67表达呈现阳性(45.0%),两组阳性率差异有统计学意义(P<0.05);对照组无1p/19q缺失,实验组有13例(41.9%)1p/19q联合缺失,其中1p单独缺失2例(6.4%),19q单独缺失1例(3.2%)。Ki-67蛋白表达与1p/19q联合缺失显著相关(r=0.403,P<0.05)。结论 Ki-67蛋白表达是预测少突胶质瘤化疗敏感性的潜在分子标记物。     

胶质瘤1号和19号染色体多体与临床病理影像学因素的相关性

目的 研究与胶质瘤1号和19号染色体多体相关的临床、病理和影像学因素.方法 254例胶质瘤患者,应用荧光原位杂交法(FISH)检测肿瘤组织中染色体1p/19q缺失状态和染色体多体比率.染色体多体定义为肿瘤组织中30％以上的肿瘤细胞含有两个以上的1q和19p荧光信号.患者年龄、性别、肿瘤病理类型、级别和部位通过回顾分析病历和影像资料获得.通过单变量和多变量分析确定与1号和19号染色体相关的因素.结果 254例中,68(26.8％)例同时为1号和19号染色体多体,1号和19号染色体多体显著相关(P＜0.000 001).单变量和多变量分析显示1号和19号染色体多体与年龄小和1p/19q联合缺失相关,与性别、肿瘤病理类型、级别、肿瘤部位无关.结论 1号和19号染色体多体共存于胶质瘤中.1号和19号染色体多体不能用于预测病理类型和级别,年龄小和1p/19q联合缺失是与染色体多体相关的独立因素,提示染色体多体对良好预后可能存在预测作用.     

联合IDH1、1p/19q和ATRX对胶质瘤分子分型的临床意义

目的探讨联合异柠檬酸脱氢酶1（IDH1）、1号染色体短臂和19号染色体长臂（1p/19q）和α-地中海贫血/智力低下综合征X染色体连锁基因（ATRX）对胶质瘤分子分型的临床意义。方法回顾经术后病理证实的104例胶质瘤患者的临床及组织病理资料,采用免疫组织化学法对其存档病理组织蜡块进行IDH1突变、ATRX表达缺失检测,采用荧光原位杂交法检测1p/19q缺失情况,通过门诊复查和电话等方式随访患者的术后生存情况,联合IDH1、1p/19q和ATRX将胶质瘤患者进行分子分型（A型:IDH1突变,1p/19q缺失;B型:IDH1突变,1p/19q完整;C型:IDH1野生,ATRX突变;D型:IDH1野生,ATRX野生）,并分析不同分子分型患者临床病理特征及预后间的差异。结果联合分子分型与胶质瘤患者的年龄、组织学分级、卡氏功能状态（KPS）评分与病理分型均有关（P<0.05）,其中与病理学分型的相关性最大,而与性别、肿瘤部位和肿瘤大小无关（P>0.05）。Kaplan-Meier生存分析结果显示,4组分子分型的胶质瘤患者术后总生存期差异显著（Log-Rank:χ2=31.631,P<0.001）,其中B型患者的中位生存期最长,D型患者中位生存期最短。COX风险回归分析显示,胶质瘤患者的预后与患者年龄、组织学分级、KPS评分、病理分型、IDH1、ATRX、联合分子分型和治疗方案均独立相关（P<0.05）,而与患者性别、肿瘤部位、大小和1p/19q缺失无关（P>0.05）。结论联合IDH1突变、1p/19q杂合缺失和ATRX突变对胶质瘤患者进行分子分型,有助于临床诊断,还可能为患者术后生存情况提供参考。     

1p/19q、MGMT启动子甲基化和VEGF对不同级别胶质瘤患者的影响

目的 探讨IDH1、1p/19q、MGMT启动子甲基化、VEGF对不同级别胶质瘤患者的影响.方法 筛选了星形细胞瘤和少突胶质细胞肿瘤及胶质母细胞瘤患者30例,其中高、低级别各15例(低级别组:A组;高级别组:B组).记录患者的一般情况、出现肿瘤相关功能缺失的临床症状、体征、术前术后影像学特点;记录的患者分子病理IDH1...     

少突胶质细胞瘤染色体1p/19q, 6-氧-甲基鸟嘌呤-DNA甲基转移酶甲基化，p53蛋白表达情况及病人预后关系的研究

目的：本实验旨在研究提高少突胶质细胞瘤的临床诊断和病人预后的相关指标。前期试验证实了少突胶质细胞瘤有很高的比例发生染色体1p/19q缺失。因此,我们进一步检测染色体1p/19q缺失,与6-氧-甲基鸟嘌呤-DNA甲基转移酶(MGMT)基因甲基化,p53蛋白表达之间的关系,及与病人预后关系。方法：用PCR-微卫星技术检测161胶质瘤染色体1p/19q缺失情况;巢式甲基化特异性PCR法检测MGMT基因甲基化情况;免疫组织化学法检测MGMT和p53蛋白表达情况。结果：少突胶质细胞瘤和星形细胞源性胶质瘤都易发生MGMT基因的甲基化。但甲基化特异性PCR法检测MGMT基因甲基化结果与免疫组织化学法检测MGMT蛋白表达并不完全相符。在间变性少突胶质细胞瘤中,未发生1p/19q缺失的病例常伴有p53蛋白的异常表达。在低级别少突胶质细胞瘤中,发生1p/19q缺失的病例常伴有MGMT基因的甲基化。发生1p/19q缺失或p53蛋白表达阴性的少突胶质细胞瘤,病人预后较好。但是MGMT基因有无甲基化与病人的预后无关。结论：检测少突胶质细胞瘤染色体1p/19q和p53蛋白表达情况可能提高临床病理诊断及提示病人预后。     

脂肪细胞增强子结合蛋白1在胶质瘤中高表达及与患者预后的关系

目的:探讨脂肪细胞增强子结合蛋白1(AEBP1)在胶质瘤组织中的表达情况及其与胶质瘤患者预后的关系。方法:通过挖掘肿瘤基因组图谱数据库(TCGA)和中国脑胶质瘤基因谱数据库(CGGA)数据,分析AEBP1在胶质瘤组织及正常脑组织中的表达情况以及其表达差异与胶质瘤分级、IDH突变、1 p/19 q联合缺失的关系。通过实时定量PCR及Western blot分别检测临床胶质瘤标本及正常脑组织中AEBP1 mRNA及蛋白表达水平。利用GEPIA网站对TCGA生成的生存期数据进行分析,绘制无病生存曲线及总生存期曲线。结果:TCGA及CGGA数据库数据分析均显示AEBP1在胶质瘤组织中相对于正常脑组织高表达,且随着胶质瘤分级的增高表达也逐级增高。AEBP1在IDH野生型、非1 p/19 q联合缺失型中表达分别高于IDH突变型及1 p/19 q联合缺失型。临床标本同样证实AEBP1 mRNA及蛋白在胶质瘤中高表达。生存期分析显示AEBP1高表达患者无病生存期及总生存期均较短,差异具有统计学意义(P<0.05)。结论:AEBP1在胶质瘤组织中高表达并预示患者预后不良。     

人脑胶质瘤中p16基因的表达

目的 探讨胶质瘤中p16基因的缺失及免疫组化情况.方法 利用、PCR技术和免疫组化检测43例胶质瘤p16基因的缺失.结果 PCR组:12例脑星形细胞瘤p16E1和p16E2均缺失,占28名(12/43),2例仅缺失p16E1,占5%(2/43);p16E基因缺失多发生在Ⅲ-Ⅳ级脑星形细胞瘤中(Ⅲ级6/16,Ⅳ级8/14),Ⅰ、Ⅱ级与Ⅲ、Ⅳ级比较有显著性差异(P<0.05).免疫组化组:p16表达缺失率为60.5%.P16表达缺失在Ⅲ级(43.6%)、Ⅳ级(85.7%)脑胶质瘤中显著高于Ⅰ-Ⅱ级(22.2%)(P<0.05).结论 p16基因失活可能与脑星形细胞瘤的分化程度有关,p16基因缺失是p16基因失活的主要机制.     

脑神经节细胞胶质瘤临床病理研究及文献复习

目的 复习文献,对1例较为罕见的脑神经节细胞胶质瘤的临床病理学特点、免疫组化染色结果及患者的预后等作一初步探讨.方法 标本采用常规石蜡切片,HE染色,先镜观察.只对石蜡切片采用EnVision免疫组化二步法染色.结果 患者男性,42岁,肿瘤位于左额颞顶叶,大部分呈囊性,镜下见肿瘤内有多量神经节细胞及星形胶质细胞和少量少枝胶质细胞.肿瘤内神经节细胞表达Syn、CgA、S-100及NSE阳性,胶质细胞表达GFAP阳性.结论 本文对肿瘤的临床、病理形态学特点、病理诊断标准及患者的预后等做了探讨.     

多形性脑胶质母细胞瘤中1p/19q杂合性缺失的表达及其意义

目的探讨多形性胶质母细胞瘤（GBM）中1p/19q杂合性缺失（1p/19q LOH）的表达类型及其意义。方法通过荧光原位杂交法检测29例GBM、17例间变性星形细胞瘤（AA）和12例少突星形细胞瘤（OA）中1p/19q LOH的表达及其差异。结果 GBM、AA和OA中1p/19q LOH的发生率分别为24.1%、35.3%和75.0%,其中同时发生1p和19q LOH的比率分别为13.8%、23.5%和66.7%,GBM中1p/19q LOH的发生率明显低于AA和OA（P＜0.05）。GBM的VEGF阳性表达率和Ki-67增殖指数分别为75.9%和87.66±16.21,AA为47.1%和49.57±4.73,OA为41.7%和41.62±4.25,GBM与后两者比较,均有统计学差异（均P＜0.05）。1p/19q LOH的发生率与VEGF阳性表达和Ki-67增殖指数呈负相关（均P＜0.05）。结论1p/19q LOH的表达可能有助于评估GBM患者对烷化剂类药物治疗的敏感性及其预后。     

两种类型染色体1p缺失在胶质瘤中的反向意义

1号染色体(1p)短臂缺失被认为是神经胶质瘤的有利预后因素。对108例胶质瘤的高密度阵列比较基因组杂交分析表明存在2种类型的1p缺失。1p全部半合子缺失与19q缺失和少突神经胶质瘤表型密切相关;而1p部分缺失主要见于星形细胞瘤,与19q缺失无关。尽管前者预示着更长的总存活期和无     

Correlation of histomorphologic prognostic markers and proliferative index with loss of heterozygosity 1p/19q and MGMT status in diffusely infiltrating gliomas

Background

Loss of heterozygosity (LOH)1p/19q, and epigenetic silencing of O⁶-methylguanine-DNAmethyltransferase (MGMT) gene, displayed promising role as predictive and prognostic markers in brain tumours. The present study correlated both with conventional histomorphologic prognostic markers and proliferative index in diffusely infiltrating gliomas (DIG).

Methods

Tissues from 45 patients were evaluated for LOH1p/19q using polymerase chain reaction based microsatellite analysis; and for MGMT using immunohistochemistry. Results were correlated with age, histologic type, WHO grade, and proliferation index.

Results

Mean MIB-1 LI showed significant correlation with tumour grade. MGMT-staining in grade II and IV tumours were 31.1% and 16.8%, respectively, while in DA and GBM it was 88.2% and 19.0%, respectively, which were statistically significant. Sixteen cases showed LOH 1p and/or 19q of which 10 (5 oligodendroglial, 3 GBM, AA, DA) had combined LOH; while three each showed 1p (all GBM) and 19q (2 DA and GBM) loss. In the MIB-1LI ≤ 5% and >5% groups LOH 1p and/or 19q was encountered in 6 and 10 cases, respectively. A significant inverse association was noted between LOH with MGMT.

Conclusions

LOH1p/19q and MGMT shows good correlation with conventional histomorphologic and proliferation markers, and should constitute part of the optimal diagnostic workup of DIG.     

Correlation of histomorphologic prognostic markers and proliferative index with loss of heterozygosity 1p/19q and MGMT status in diffusely infiltrating gliomas

Background

Loss of heterozygosity (LOH)1p/19q, and epigenetic silencing of O⁶-methylguanine-DNAmethyltransferase (MGMT) gene, displayed promising role as predictive and prognostic markers in brain tumours. The present study correlated both with conventional histomorphologic prognostic markers and proliferative index in diffusely infiltrating gliomas (DIG).

Methods

Tissues from 45 patients were evaluated for LOH1p/19q using polymerase chain reaction based microsatellite analysis; and for MGMT using immunohistochemistry. Results were correlated with age, histologic type, WHO grade, and proliferation index.

Results

Mean MIB-1 LI showed significant correlation with tumour grade. MGMT-staining in grade II and IV tumours were 31.1% and 16.8%, respectively, while in DA and GBM it was 88.2% and 19.0%, respectively, which were statistically significant. Sixteen cases showed LOH 1p and/or 19q of which 10 (5 oligodendroglial, 3 GBM, AA, DA) had combined LOH; while three each showed 1p (all GBM) and 19q (2 DA and GBM) loss. In the MIB-1LI ≤ 5% and >5% groups LOH 1p and/or 19q was encountered in 6 and 10 cases, respectively. A significant inverse association was noted between LOH with MGMT.

Conclusions

LOH1p/19q and MGMT shows good correlation with conventional histomorphologic and proliferation markers, and should constitute part of the optimal diagnostic workup of DIG.     

p53 mutation, EGFR gene amplification and loss of heterozygosity on chromosome 10, 17 p in human gliomas

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Analysis of comparative genomic hybridization and loss of heterozygosity in 43 primary gastric carcinomas

Objective To investigate common chromosomal changes and the LOH frequency of microsatellite loci in primary gastric cancer samples in order to locate the deleted regions in which human gastric cancer related genes might exist.Methods Comparative genomic hybridization (CGH) was used to define global chromosomal aberrations in 43 primary gastric tumors. Based on the results of CGH, analysis of loss of heterozygosity (LOH) was performed in chromosome 19 in which the loss was first discovered in the gastric cancers. The PCR-based approach was used to investigate 22 loci, which are spaced at 1.1 -10. 9 cM intervals throughout chromosome 19. The amplified PCR fragments were subjected to electrophoresis in PAGE gel and analyzed with GenescanTM and GenotyperTM.Results CGH analysis revealed gains in chromosome 3p(8/43), 8q(8/43), 20 [20 (9/43), 20p(7/43), 20q(4/43)], 12q(16/43), 13q(12/43) and losses in 19 [19 (15/43)], 7 [17 (8/43),17p (10/43)], 16 (10/43) and lp (11/43). Among the 43 evaluated samples, the most frequent LOH was detected at locus D19S571 (27. 81% ).Conclusions The tumorigenesis of gastric cancer includes several chromosomal changes. The aberration of chromosome 19 was the first common change founded in gastric cancer. The region near the D19S571 miclht harbor potential clenes related to the tumoriQenesis of Qastric cancer.     

食管不典型增生和早期食管鳞癌全基因组的变化特征分析

目的 应用SNP芯片检测食管不典型增生和早期食管鳞癌全基因组的DNA拷贝数异常和杂合性缺失(LOH),探讨它们的变化特征.方法 应用Affymetrix GeneChip Human Mapping250K NspArray芯片检测1例原发性食管重度不典型增生和4例原发性早期食管鳞癌病变组织和配对正常食管组织的DNA拷贝数的变化和LOH.结果 食管重度不典型增生只有极少数的DNA片段发生扩增,未发生缺失或LOH.早期食管鳞癌发生DNA扩增的染色体有1p、1q、2p、2q、3q、4q、5p、6p、6q、7q、8q、11p、11q、12p、12q、14q、17q、18p、19q、20q、22q和X;发生DNA缺失的染色体有1p、2q、3p、3q、4p、4q、8p、9p、9q、10q、11p、13q、16p、18q、19p、19q和22q;发生LOH的染色体有3q和9q.其中1p、19q、4q、11p等染色体发生扩增和3q、11p、2q、16p等染色体发生缺失罕见报道.结论 食管重度不典型增生DNA的变异不明显;早期食管鳞癌DNA发生明显的扩增和缺失,但很少发生LOH.250K SNP芯片能够有效地检测食管不典型增生和早期食管鳞癌全基因组范围DNA拷贝数的变化及LOH,分辨率高,定位精确,这些结果为进一步定位筛选和克隆与早期食管鳞癌相关的基因提供了重要的理论信息.     

19号染色体微卫星杂合性缺失与原发性胃癌的临床关系

目的:研究19号染色体短臂微卫星不稳定性(microsatellite instability,MSI)及杂合性缺失(loss of heterozygosity,LOH)与胃癌临床病理特征之间的关系,探讨19号染色体短臂微卫星MSI和LOH的主要临床意义.方法:采用PCR-SSLP-银染方法扩增79例原发性胃癌及正常组织标本中19号染色体短臂不同位置的7个点,PCR产物经聚丙烯酰胺凝胶电泳分离,运用Genescan软件和Genotyper软件分析MSI和LOH,然后进一步分析微卫星LOH与原发性胃癌的临床关系.结果:79例胃癌患者中,至少有1种微卫星发生LOH,其频率为31.18%(27/79),在所有微卫星中,D19S591和D19S565的LOH发生率分别为60.32%(38/63)和48.15%(26/54),高于其他微卫星的LOH.LOH高频率与原发性胃癌的临床分期及远处转移相关,且随着恶性程度增加LOH频率也增加(P＜0.05),而MSI与胃癌临床病理之间相关性不大.结论:19p高频率的LOH与原发性胃癌的临床分期和远处转移相关,且LOH高频率提示在该区域可能存在肿瘤抑癌基因,其与胃癌的发生及进展相关.     

原发性肝癌患者17号和16号染色体等位基因杂合性丢失的研究

目的 研究肝癌１７号和１６号染色体特定区域等位基因杂合性丢失（ＬＯＨ）发生情况,并探讨ＬＯＨ与肝癌的临床病理和乙型、丙型肝炎病毒感染的关系。方法 用聚合酶链反应、微卫星多态性分析技术检测１７号和１６号染色体ＬＯＨ。结果 １７号染色体６个位点在至少一个位点发生ＬＯＨ的有３１例（８２％）,其中Ｄ１７Ｓ５２０（１７ｐ１２－１３．３）和ＴＰ５３（１７ｐ１３．１）位点ＬＯＨ频率大于５０％;１７ｑ所选４个位点     

Presence of allelic loss and PTEN mutations in malignant gliomas from Malay patients

Loss of heterozygosity (LOH) on several loci and mutations on PTEN tumor suppressor gene (10q23.3) occur frequently in sporadic gliomas. We have performed polymerase chain reaction (PCR)-LOH analysis using microsatellite markers and single-stranded conformational polymorphism (SSCP) analysis to determine the incidence of allelic losses on chromosome 10q, 9p, 17p and 13q and mutations of exons 5, 6 and 8 of the PTEN gene in malignant gliomas. Twelve of 23 (52.2%) malignant glioma cases showed allelic losses whereas 7 of 23, (30.4%) samples showed aberrant band patterns and mutations of the PTEN gene. Four of these cases showed LOH on 10q23 and mutations of the PTEN gene. The data on LOH indicated the involvement of different genes in gliomagenesis whereas mutations of the PTEN gene indicated the role of PTEN tumor suppressor gene in the progression of glioma in Malay population.     

Chromosome 1p/19q status combined with expression of protein improves the diagnostic and prognostic evaluation of oligodendrogliomas

Background Our previous study confirmed that oligodendrogliomas had higher frequency of chromosome 1p/19q deletion. In order to improve the diagnostic criteria and to predict the prognosis of oligodendroglioma patients, the status of chromosome 1 p/19q deletion, the methylation of O6-methylguanine-DNA methyltransferase (MGMT), and the expression of p53 protein were evaluated and investigated in relation to patients' outcomes.Methods Methylation of MGMT in 73 cases was analyzed by nested methylation-specific PCR (MSP). The levels of MGMT and p53 protein were tested with immunohistochemistry. Pearson's chi-square test and Fisher's exact test were used. Multivariate and Kaplan-Meier analysis were performed to determine patients' outcomes.Results Both oligodendrogliomas and astrocytic gliomas exhibited frequent methylation of MGMT. However, the results of MSP did not completely correspond to that of the immunohistochemical staining for MGMT. The expression of p53 protein was more frequently observed in patients without a 1 p or 19q deletion in anaplastic oligodendrogliomas (=0.032,0.025). In low-grade oligodendrogliomas, methylation of MGMT was more frequent in patients with 1 p/19q deletion than in patients with 1p/19q intact (P=0.038). Patients with oligodendrogliomas with 1p/19q loss of heterozygosity and p53-negative showed a longer progression-free survival.Conclusion Detection of chromosome 1p/19q status combined with p53 protein immunohistochemistry might be beneficial to improve the pathological diagnosis and to determine the prognosis of patients with oligodendrogliomas.     

Genome-wide allelotype study of primary glioblastoma multiforme

Objective To investigate the molecular genetic pathogenesis of primary glioblastoma multiforme (GBM) and identify which chromosomes or chromosomal regions of the entire genome may harbor tumor suppressor genes (TSGs) associated with GBM.Methods A high-resolution allelotype study of 21 cases of primary GBM was performed by PCR-based loss of heterozygosity （LOH）analysis. Three hundred and eighty-two fluorescent dye-labeled microsatellite markers covering all 22 autosomes were applied. The mean genetic distance between two flanking markers was about 10 cM.Results LOH was observed on all 39 nonacrocentric autosomal arms examined in this study. The LOH frequencies of 10q, 10p, 9p, 17p and 13q were the highest (&gt;50%). Furthermore, high LOH frequencies were detected in the regions containing known TSGs including PTEN, DMBT1, p16, p15, p53 and RB; the LOH frequencies on 14q, 3q, 22q, 11p, 9q, 19q were also high (&gt;40.5%). Our study observed the following commonly deleted regions: 9p22-23, 10p12.2-14, 10q21.3, 13q12.1-14.1, 13q14.3-31, 17p11.2-12, 17p13, 3q25.2-26.2, 11p12-13, 14q13-31, 14q32.1, 14q11.1-13, 22q13.3, 4q35, 4q31.1-31.2, 6q27 and 6q21-23.3. Conclusions The molecular pathogenesis of GBM is very complicated and associated with a variety of genetic abnormalities on many chromosomal arms. The most closely related chromosomal arms to the pathogenesis of GBM are 10q, 10p, 9p, 17p and 13q. Besides the well-known TSGs including PTEN, DMBT1, p16, p15, p53 and RB, multiple unknown TSGs associated with GBM may be present on the commonly deleted regions detected in the present study.