NDUFA4L2在胶质瘤组织中的表达及意义

［摘要］　目的　探讨乳酸脱氢酶A的α复合体4-L2（NDUFA4L2）在胶质瘤中的表达情况及临床意义。方法　利用癌症基因组图谱（TCGA）、中国胶质瘤基因组图谱（CGGA）数据库分析NDUFA4L2在胶质瘤中的mRNA表达情况,分析其与患者临床病理分级和预后的关系。进一步收集不同级别的胶质瘤组织标本行免疫组化检测,验证NDUFA4L2在不同级别胶质瘤中蛋白表达的差异。收集62例胶质瘤患者的组织标本和随访资料,采用RT-PCR法检测NDUFA4L2在胶质瘤中的表达并分析其与预后的关系。结果　通过分析TCGA、CGGA数据库,发现胶质瘤组织中NDUFA4L2的表达随着肿瘤分级的增高而增高(P<0.05),高表达NDUFA4L2预示着患者生存期较短。在临床标本验证中的结果与上述生物信息学分析相一致。结论　NDUFA4L2在胶质瘤组织中的表达与病理分期和预后有关,可能是胶质瘤的一个潜在肿瘤标志物。     

磁共振扩散峰度成像与脑星形细胞瘤GFAP表达的相关性研究

目的 应用磁共振扩散峰度成像（DKI）初步探讨脑星形细胞瘤的DKI 参数值与术后病理GFAP 表达的相关性.方法 收集经手术病理证实的脑星形细胞瘤66 例,其中高级别（WHO Ⅲ级-Ⅳ级）34 例,低级别（WHO Ⅰ级-Ⅱ级）32 例,术前行常规磁共振平扫增强及DKI 扫描,通过工作站对图像进行后处理,测定肿瘤实质区部分各向异性分数（FA）值及平均扩散峰度（MK）值,采用两样本t 检验比较两组间DKI 各参数值及GFAP 表达水平差异,采用Spearman 秩相关分析DKI 各参数值与GFAP 表达的相关性.结果 高级别组脑星形细胞瘤实质区MK 值明显高于低级别组（P 〈0.05）;高级别组脑星形细胞瘤实质区FA 值与低级别组差异无统计学意义（P = 0.331）;高级别组脑星形细胞瘤实质区GFAP 表达显著低于低级别组（P 〈0.05）;MK 值与GFAP 表达呈显著负相关（r = -0.836;P = 0.03）;FA 值与GFAP（r = -0.562;P = 0.05）表达无相关性.结论 DKI 参数MK 值与脑星形细胞瘤GFAP 表达显著相关.应用DKI 在一定程度上可以补充常规磁共振,间接评估肿瘤细胞的分化程度、恶性程度、浸润转移等生物学行为,有助于脑星形细胞瘤的个体诊断及治疗.     

平均峰度在脑星形细胞瘤分级中的应用价值

目的：探讨在扩散峰度成像（DKI）中的平均峰度（MK）值对不同级别脑星形细胞瘤分级的应用价值。方法回顾性分析2012年3月-2013年12月山西医科大学第一医院经病理证实的30例脑星形细胞瘤患者,术前行头颅MRI平扫、增强扫描和DKI检查。测定WHOⅡ、Ⅲ和Ⅳ级星形细胞瘤不同微观结构的MK值,均与对侧对应的正常脑白质MK值进行标准化处理,标准化处理所得的MK值,方差齐性检验后,采用LSD法进行组间比较;在同一级别肿瘤组内, MK值采用LSD法进行组内比较。结果在高、低级别脑星形细胞瘤组间肿瘤实质区MK值有统计学意义（P〈0.05）。WHOⅢ级肿瘤实质与远侧比较、囊变区与近侧水肿带比较, MK值差异无统计学意义（P〉0.05）;WHOⅣ级各微观结构两两比较,其MK值差异均有统计学意义（P〈0.05）。结论肿瘤实质部分MK值提示有助于高低级别脑星形细胞瘤分级,但尚不能提示有助于WHOⅢ级和Ⅳ级脑星形细胞瘤分级,WHO Ⅳ脑星形细胞瘤的囊变区与近侧水肿带差异比 WHOⅢ级肿瘤明显,有助于 WHOⅢ级和Ⅳ级脑星形细胞瘤鉴别。     

ADAMTS7基因表达水平对神经胶质瘤患者预后作用及临床病理特征的关联性探讨

目的探讨血小板反应蛋白提取整合素金属肽酶7(ADAMTS7)基因表达水平对神经胶质瘤患者预后作用及临床病理特征的关联性。方法在中国胶质瘤基因组图谱(CGGA)数据库(http://www.cgga.org.cn)下载mRNAseq 693和mRNAseq 325两个转录组测序数据集,以mRNAseq 693作为探索数据集,mRNAseq 325作为验证数据集。提取数据集病例的ADAMTS7基因表达量,以ADAMTS7的中位值作为截断阈值将患者分为高表达组和低表达组。分析ADAMTS7基因表达水平与患者临床病理特征的关联性及对患者生存预后的影响,并以美国癌症基因组图谱数据库(TCGA)胶质瘤队列作为外部数据集再次验证ADAMTS7基因表达水平与胶质瘤患者预后的关系。结果无论是在探索数据集还是在验证数据集中,ADAMTS7基因高表达组与低表达组在发生状态、组织学类型、WHO分级、化疗状态、异柠檬酸脱氢酶(IDH)突变状态和1号染色体短臂和19号染色体长臂(1p19q)共缺失状态方面比较差异均有统计学意义(P 0.05)。对于探索数据集,多因素Cox回归分析结果显示,较高的ADAMTS7基因表达水平、疾病复发、WHO分级为Ⅳ级和年龄 43岁是影响患者术后生存预后的危险因素,而接受化疗、IDH突变和1p19q共缺失是影响患者术后生存预后的保护因素(P 0.05)。对于验证数据集,多因素Cox回归分析结果显示,较高的ADAMTS7基因表达水平、疾病复发、WHO分级为Ⅳ级是影响患者术后生存预后的危险因素,而组织学类型为多形胶质母细胞瘤(GBM)、接受化疗和1p19q共缺失是影响患者术后生存预后的保护因素(P 0.05)。Kaplan-Meier生存分析结果显示,无论是对于探索数据集还是验证数据集,ADAMTS7基因低表达组的术后生存预后均显著优于高表达组(P 0.001)。以TCGA胶质瘤数据作为外部数据集验证,结果显示,ADAMTS7基因高表达组的总生存期(OS)和无进展间期(PFI)优于低表达组,差异有统计学意义(P 0.001)。结论 ADAMTS7基因表达水平可作为评估神经胶质瘤患者预后的预测因子,具有潜在的临床应用价值。     

转化生长因子-β受体Ⅲ在胶质瘤组织中的表达及意义

目的探讨转化生长因子-β受体Ⅲ(TβRⅢ)在胶质瘤发生发展中的作用。方法采用免疫组化法检测TβRⅢ在3例正常脑组织、13例低级别(Ⅰ、Ⅱ级)胶质瘤和27例高级别(Ⅲ、Ⅳ级)胶质瘤中的表达,Western Blot法进一步验证各级别不同病理类型中TβRⅢ表达。结果正常脑组织中TβRⅢ阳性表达率为100%(3/3),低级别胶质瘤阳性表达率为84.6%(11/13),高级别胶质瘤阳性表达率为48.1%(13/27),高、低级别间比较,P<0.05;Ⅱ、Ⅲ级肿瘤中,星形细胞瘤和含少突成分的胶质瘤间TβRⅢ的表达无显著差异。结论高级别胶质瘤中TβRⅢ表达降低;TβRⅢ可能参与了胶质瘤的恶变过程,其低表达可促进肿瘤的侵袭和迁移。     

血浆IGFBP-2在高级别脑胶质瘤手术及放化疗前后的变化

目的探讨动态监测脑胶质瘤患者血浆胰岛素样生长因子结合蛋白2（IGFBP-2）在病情判定、预测复发中的临床意义。方法用ELISA法检测健康体检者及不同级别胶质瘤患者在术前、综合治疗后及复发前后血浆IG-FBP-2水平。随访其中55例高级别胶质瘤患者2.5 a,分析术前血浆IGFBP-2水平与其无瘤生存期的关系。结果①血浆IGFBP-2水平随胶质瘤级别的升高而增加（P〈0.01）;②高级别胶质细胞瘤患者经过手术、放疗、化疗综合治疗后未复发者血浆IGFBP-2下降（P〈0.05）,而复发后升高（P=0.01）;③高级别脑胶质瘤患者血浆中IGFBP-2水平〈600 ng/ml的无瘤生存期长于〉600 ng/ml者（P〈0.05）。结论血浆IGFBP-2的动态监测对判定胶质瘤患者病变恶性程度及预测高级别胶质瘤的复发有一定临床意义。     

星形胶质细胞瘤中HSP70的表达及生物学活性

目的探讨星形胶质细胞瘤中热休克蛋白(HSP70)表达水平与肿瘤恶性程度的关系及其与胶质细胞瘤生物学行为的关系。方法采用HSP70单克隆抗体对62例星形胶质瘤进行免疫组化染色,检测HSP70的表达水平及其与胶质瘤病理分级的关系,随访观察患者5年生存率,比较不同HSP70表达水平下患者的生存率差异。结果 62例星形胶质细胞瘤患者中,21例HSP70表达增高,与正常脑组织有统计学差异(P0.05);HSP70表达情况与性别、年龄无关(P0.05);HSP70的表达水平与胶质瘤病理分级呈正相关(r=0.685,P0.01);HSP70阳性组5年生存率明显低于HSP70阴性组(χ2=4.742,P0.05)。结论 HSP70与胶质瘤的生物学行为和分化程度有关,HSP70的检测会对星形胶质细胞瘤的诊断与靶向治疗提供实验依据。     

胶质瘤IDH1基因变异与Ki-67、微血管密度表达的相关性

目的探讨胶质瘤IDH1基因变异对组织中抗原Ki-67及微血管密度(MVD)表达的影响及IDH1基因变异延长患者生存期的机制。方法运用SP法分别进行Ki-67、CD34的免疫组织化学染色检测胶质瘤IDH1变异组及未变异组抗原Ki-67标记指数(LI)及MVD计数。结果变异组Ki-67蛋白表达较未变异组降低(t=2.134,P=0.039),变异组与未变异组Ki-67 LI分别为(14.44%±8.23%)和(21.19%±11.49%);变异组MVD较未变异组减低(t=2.328,P=0.025),其MVD分别为31.11±13.47和40.54±12.11;低级别胶质瘤Ki-67及MVD表达较高级别胶质瘤低(t=9.138,P<0.001;t=-8.369,P<0.001),低级别胶质瘤(WHOⅠ级、Ⅱ级)和高级别胶质瘤(WHOⅢ级、Ⅳ级)Ki-67表达分别是8.36%±2.84%和25.55%±7.53%,其MVD分别为23.94±8.03和45.54±8.19,两者表达随胶质瘤级别增高而增高。结论胶质瘤IDH1基因变异对肿瘤细胞的增殖有影响,IDH1基因变异影响肿瘤的能量代谢,而能量代谢受阻降低细胞的增殖活性,使大量细胞处于休眠期,导致新生血管内皮细胞分裂缓慢,新生肿瘤血管数量减低,恶变和转移受到抑制,为IDH1基因变异延长患者的生存期的机制提供新的理论依据。     

星形胶质细胞瘤的PTEN、HIF-1α、VEGF表达及临床意义

目的探讨星形胶质细胞瘤中的与张力蛋白和辅助蛋白同源、第10号染色体丢失的磷酸基因（PTEN）缺失情况，PTEN与缺氧诱导因子-1α（HIF—1α）、血管内皮生长因子（VEGF）以圾与某些临床因素的关系。方法用逆转录一聚合酶链反应（RT—PCR）检测PTEN基因缺失；应用免疫组化法检测肿瘤中PTEN、HIF-1α、VEGF蛋白表达情况。结果星形胶质细胞瘤的PTEN缺失率为12．5％（4／32），均为Ⅳ级。PTEN、HIF-1α、VEGF蛋白在不同恶性级别星形胶质瘤细胞中的阳性表达率均具有差异性，与肿瘤恶性程度呈相关。PTEN与HIF-1α、VEGF表达呈负相关，HIF-1α和VEGF表达呈正相关。结论PTEN通过对HIF—1α、VEGF的作用参与了肿瘤的恶性进展过程，HIF—1α是VEGF表达调控因子之一。三者相互作用。对星形胶质细胞瘤的进展，尤其是对肿瘤血管新生起有重要作用。     

TGF-β1、Smad7在老年星形细胞瘤患者术后组织中的表达及其相关性

目的探讨转化生长因子(TGF)-β1、Smad7在老年星形细胞瘤患者术后组织中的表达及其相关性。方法接受手术治疗的老年星形细胞瘤患者25例为研究组。取术中切除的星形细胞瘤组织标本,分为低级别(Ⅰ~Ⅱ级)11例,高级别(Ⅲ~Ⅳ级)14例;另收集本院行内减压术切除的额颞叶正常脑组织6例为对照组。免疫组化染色检测TGF-β1、Smad7在星形细胞瘤及正常脑组织中的表达,RT-PCR检测TGF-β1、Smad7mRNA表达水平。结果 TGF-β1、Smad7主要表达于正常脑组织和星形细胞瘤组织的细胞质,为黄色或棕黄色颗粒,细胞膜和细胞核均无明显阳性表达;各组间TGF-β1、Smad7表达水平差异均有显著统计学意义(P<0.01),TGF-β1、Smad7的阳性表达率由高到低依次为:高级别星形细胞瘤、低级别星形细胞瘤、正常脑组织,TGF-β1与Smad7之间呈明显正相关。正常脑组织、低级别星形细胞瘤、高级别星形细胞瘤中TGF-β1 mRNA和Smad7mRNA表达量逐渐增加,两两比较差异均有统计学意义(P<0.05)。结论 TGF-β1、Smad7表达与老年星形细胞瘤的病理分级呈正相关,TGF-β1/Smad信号通路参与了星形细胞瘤的异常增殖。     

Tumor-associated macrophages based signaling pathway analysis and hub genes identification in glioma

Tumor-associated macrophages (TAMs) play a crucial role in the immune response to many malignancies, but the signaling pathways by which the glioma microenvironment cross-talk with TAMs are poorly understood. The aim of this study was to uncover the potential signaling pathways of the regulation of TAMs and identify candidate targets for therapeutic intervention of glioma through bioinformatics analysis.Chinese Glioma Genome Atlas (CGGA) and The Cancer Genome Atlas (TCGA) datasets were used to download RNA-Seq data and microarray data of human glioma specimen. Differentially expressed genes (DEGs) between CD68-high samples and CD68-low samples were sorted. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the DEGs was conducted. Protein-protein interaction (PPI) network were formed to identify the hub genes.The prognostic value of TAMs in glioma patients was confirmed. A total of 477 specific DEGs were sorted. The signaling pathway was identified in pathway enrichment and the DEGs showed prominent representations of immune response networks in glioma. The hub genes including C3, IL6, ITGB2, PTAFR, TIMP1 and VAMP8 were identified form the PPI network and they were all correlated positively with the expression of CD68 and showed the excellent prognostic value in glioma patients.TAMs can be used as a good prognostic indicator in glioma patients. By analyzing comprehensive bioinformatics data, we uncovered the underlying signaling pathway of the DEGs between glioma patients with high and low expression level of CD68. Furthermore, the 6 hub genes identified were closely associated with TAMs in glioma microenvironment and need further investigation.     

EFNA1 is a potential key gene that correlates with immune infiltration in low-grade glioma

EFNA1 is a key gene that is associated with the pathogenesis of several human cancers. However, the prognostic role of EFNA1 in many cancers and the relationship between EFNA1 and tumor-infiltrating lymphocytes in different cancers remain unclear.The expression levels of EFNA1 in 33 types of cancer in the TCGA (The Cancer Genome Atlas) database were collected via the UCSC Xena browser. The clinical data of LGG (low grade glioma) patients were downloaded from the TCGA database. The glioma data from the CGGA (Chinese Glioma Genome Atlas) database were also downloaded to verify the results. Kaplan–Meier and Cox regression analyses were used to investigate the prognostic value of EFNA1 in different cancers using R software. We verified the differential expression of EFNA1 in glioma and normal brain tissue via gene expression profiling interactive analysis. We evaluated the relationship between the expression level of EFNA1 and the clinicopathological features of LGG patients via the Wilcoxon signed-rank test. The immune infiltration levels were evaluated via tumor immune estimation resource (TIMER) and CIBERSORT, and the correlations between EFNA1 and immune cell levels were investigated via TIMER. Finally, we conducted gene set enrichment analysis (GSEA) to explore the potential mechanisms.Data from the TCGA database showed that EFNA1 was differentially expressed in many kinds of cancers when compared with normal tissues. Upregulated EFNA1 expression in esophageal carcinoma (ESCA), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), and LGG correlated with shorter patient overall survival (OS) times. The Cox regression analysis revealed that the expression of EFNA1 was also a risk factor for the disease-specific survival (DSS) and progression-free interval (PFI) of LGG patients. The multiple Cox regression analysis revealed that EFNA1 was an independent prognostic factor for LGG patients. In addition, EFNA1 expression was increased in the WHO grade III group and the 1p19q non-codeletion group. Moreover, EFNA1 expression was positively correlated with the levels of infiltrating CD4+ T cells, myeloid dendritic cells and neutrophils in LGG. GSEA suggested that several GO and kyoto encyclopedia of genes and genomes (KEGG) items associated with nervous system function and apoptotic pathway were significantly enriched in the EFNA1-low and EFNA1-high expression phenotypes.EFNA1 may play a pivotal role in the development of LGG and may serve as a potential marker for LGG prognosis and therapy.     

乙型肝炎病毒相关肝细胞癌核心差异表达基因生物信息学分析

目的 探索与乙型肝炎病毒（HBV）相关肝细胞癌（HCC）发生发展相关的核心基因,为进一步揭示HBV相关HCC发病机制提供参考。方法 从高通量基因表达数据库（GEO）中下载GSE55092、GSE121248两个数据集,采用R语言筛选HCC组织和癌旁组织间差异表达基因（DEGs）,并绘制可视化火山图。对DEGs基因进行基因本体论（GO）和京都基因和基因组百科全书（KEGG）富集分析,构建蛋白质相互作用（PPI）网络,并用Cytoscape 3.9.0开源平台中分子复合物检测（MCODE）和cytoHubba插件筛选核心DEGs。利用UALCAN和Kaplan Meier-plotter数据库中临床样本数据对筛选出的核心DEGs进行差异表达和生存分析验证。结果 从GSE55092数据集和GSE121248数据集中分别筛选出1 148个和686个DEGs,其中下调表达基因分别为703个和477个、上调表达基因分别为445个和209个;两个数据集共筛选出557个共同表达的DEGs,其中下调表达基因384个、上调表达基因173个。GO富集分析显示,DEGs主要参与细胞分裂、细胞增殖、氧化还原、免...     

STIL在神经胶质瘤中表达及对增殖能力的影响

目的 探讨STIL在神经胶质瘤中表达及生物学意义.方法 公用数据库TCGA在线分析STIL在神经胶质瘤中表达水平,并检测了20例神经胶质瘤临床标本中STIL表达量,利用siRNA干扰技术瞬时敲低胶质瘤细胞系中STIL的表达水平,并利用定量聚合酶链反应(qPCR)及Western印迹检测敲低效率,免疫组织化学染色(SP)...     

基于多平台数据识别胰腺导管腺癌预后相关肿瘤微环境基因

目的 探索与胰腺导管腺癌(PDAC)相关的肿瘤微环境基因表达模块,识别影响患者预后的生物学标志物和潜在的免疫治疗靶点。方法 筛选收集来自肿瘤基因组图谱(TCGA)数据库的1个包含142例PDAC患者数据集和基因表达综合(GEO)数据库的2个包含168例PDAC患者微阵列数据集(GSE2150、GSE62452)的基因表达谱数据,运用xCell网络工具对PDAC基因表达数据进行细胞类型富集分析。根据细胞富集评分中位数将TCGA的142例患者分为高分、低分2组,通过单因素生存分析确定有预后价值的细胞类型并使用GEO的数据集验证。再根据细胞类型进行基因差异表达分析及单因素生存分析确定与预后相关的差异表达基因(DEGs),并对DEGs进行功能富集分析及蛋白质-蛋白质相互作用(PPI)网络分析。同时使用GEO数据集对TCGA数据集的预后相关DEGs进行验证。最后在TISIDB数据库检索TCGA与GEO数据库的共同DEGs,并分析其与免疫系统的相关性。结果 细胞类型富集评分显示,Th1细胞和角质形成细胞在TCGA和GEO数据集中具有相同的预后价值,其高分组患者的总生存率显著低于低分组,差异均有统计学意义(P值均<0.05)。鉴定出216个预后相关DEGs,对其功能富集结果显示21个生物学过程条目中有9个与免疫过程密切相关,5条京都基因与基因组百科全书(KEGG)通路中有4条与免疫过程密切相关。通过PPI网络分析,CCR7、CD27、CD5、CXCL13、ZAP70、MS4A1和CCL19被鉴定为可能与PDAC密切相关的中枢基因。通过对GEO数据集的验证,共有15个DEGs在GEO和TCGA数据集中具有相似的预后价值。检索TISIDB数据库上述15个基因结果显示,GIMAP7与PDAC的免疫过程密切相关。结论 鉴定了216个与PDAC预后相关的肿瘤微环境基因及其7个中枢基因,并提供了CCR7、CCL19、CD27、CXCL13和GIMAP7 5个新的PDAC免疫治疗潜在靶点。     

Identification of potential biomarkers of peripheral blood mononuclear cell in hepatocellular carcinoma using bioinformatic analysis: A protocol for systematic review and meta-analysis

Background:Hepatocellular carcinoma (HCC) is the cause of an overwhelming number of cancer-related deaths across the world. Developing precise and noninvasive biomarkers is critical for diagnosing HCC. Our research was designed to explore potentially useful biomarkers of host peripheral blood mononuclear cell (PBMC) in HCC by integrating comprehensive bioinformatic analysis.Methods:Gene expression data of PBMC in both healthy individuals and patients with HCC were extracted from the Gene Expression Omnibus (GEO) to identify differentially expressed genes (DEGs). The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were applied to annotate the function of DEGs. Protein-protein interaction analysis was performed to screen the hub genes from DEGs. cBioportal database analysis was performed to assess the prognostic significance of hub genes. The Cancer Cell Line Encyclopedia (CCLE) and The Human Protein Atlas (HPA) database analyses were performed to confirm the expression levels of the hub genes in HCC cells and tissue.Results:A total of 95 DEGs were screened. Results of the GO analysis revealed that DEGs were primarily involved in platelet degranulation, cytoplasm, and protein binding. Results of the KEGG analysis indicated that DEGs were primarily enriched in focal adhesion. Five genes, namely, myosin light chain kinase (MYLK), interleukin 1 beta (IL1B), phospholipase D1 (PLD1), cortactin (CTTN), and moesin (MSN), were identified as hub genes. A search in the CCLE and HPA database showed that the expression levels of these hub genes were remarkably increased in the HCC samples. Survival analysis revealed that the overexpression of MYLK, IL1B, and PLD1 may have a significant effect on HCC survival. The aberrant high expression levels of MYLK, IL1B, and PLD1 strongly indicated worse prognosis in patients with HCC.Conclusions:The identified hub genes may be closely linked with HCC tumorigenicity and may act as potentially useful biomarkers for the prognostic prediction of HCC in PBMC samples.     

MSMEG＿6171在耻垢分枝杆菌中的表达及多组学关联分析

目的 筛选MSMEG＿6171在耻垢分枝杆菌中的可能调控通路或基因,并进行验证。方法 构建MSMEG＿6171基因过表达的耻垢分枝杆菌株和空载质粒对照菌株;对其转录组和蛋白质组进行验证并深入挖掘MSMEG＿6171过表达调控的下游信号网络;qRT-PCR验证差异表达基因。结果 成功构建了MSMGE＿6171过表达的耻垢分枝杆菌菌株(Msm::6171);转录组和蛋白质组数据的对比分析发现有38个基因在转录水平和蛋白表达水平显著上调,有31个基因表达水平显著下调;差异表达基因主要参与甘油酯代谢,核糖体和代谢通路;15个候选基因实时荧光定量PCR验证结果与转录组和蛋白质组测序分析趋势一致。结论 MSMGE＿6171调控了多个靶向基因,参与代谢和核糖体通路,为深入研究MSMGE＿6171的调控网络提供了新的认识和重要参考。