利用全基因组寡核苷酸芯片初步筛选无功能垂体腺瘤相关基因

目的探讨人无功能垂体腺瘤发生、发展中相关基因的表达与功能。方法应用人全基因组寡核苷酸芯片HG-U133Plus2.0分别检测4例无功能垂体腺瘤组织(2例促性腺激素腺瘤、2例裸细胞激素腺瘤)和正常垂体组织的基因表达谱,并对差异表达基因进行筛选和生物信息学分析。同时随机选择EDG3基因予以实时荧光定量PCR验证。结果与正常垂体组织相比,促性腺激素腺瘤中上调基因223条,下调基因678条;而裸细胞腺瘤中上调基因156条,下调基因665条;两者共同表达基因中上调基因50条,下调基因136条。共同差异表达基因的功能主要涉及分子结合、凋亡或肿瘤相关、代谢、信号传导、细胞周期、物质运输等多个生物过程。结论基因芯片技术有助于在分子水平上了解垂体腺瘤发病机制。     

泌乳素腺瘤基因表达谱的建立与分析

目的建立和分析泌乳素腺瘤的基因表达谱。方法应用人全基因组寡核苷酸芯片HG-U133A2.0检测2例泌乳素腺瘤组织和2例混合的正常垂体组织的基因表达谱,对差异表达基因进行筛选和生物信息学分析。结果成功建立正常垂体与泌乳素腺瘤的基因表达谱,筛选出与泌乳素腺瘤相关的上调基因313条,下调基因856条。这些差异基因功能聚类显示主要涉及分子结合、凋亡或肿瘤相关、代谢、信号传导、细胞周期等多个分子通路。结论基因表达谱的建立有助于分析、研究泌乳素腺瘤的分子机制。     

生长激素腺瘤中差异表达的微小RNAs及其靶基因的相关生物信息学分析

目的探讨垂体生长激素(GH)细胞腺瘤中差异表达的微小RNAs(miRNAs)及其靶基因的生物学功能和两者的调控关系。方法利用miRDB、miRwalk、Targetscan7.2及starbase数据库对差异表达的miRNAs进行靶基因预测并对靶基因进行GO、KEGG和蛋白-蛋白相互作用网(PPI)分析,随后筛选出有意义的核心靶基因,取核心靶基因和数据库的mRNAs测序结果进行对比,构建可视化的miRNAs-靶基因调控关系网。结果以蛋白相互作用程度≥20为标准,本研究得到113个上调的miRNAs核心靶基因和128个下调的miRNAs核心靶基因,进一步取交集得到13个目的核心靶基因,这些基因主要涉及的通路为泛素介导蛋白水解通路,其相对应的调控miRNAs为miR-15b、miR-365、miR-32-3p、miR-486-5p。结论本研究应用生物信息学分析工具构建了与GH细胞腺瘤密切相关的miRNAs-核心靶基因相互调控网,发掘了一些可能与GH细胞腺瘤发病机制和病理过程有关的蛋白和通路。     

利用全基因组芯片初步筛选亚类垂体腺瘤特异性相关基因

目的 探讨人类垂体腺瘤发生、发展中相关基因的表达与功能.方法 应用人全基因组寡核苷酸芯片HG-U133A2.0检测8例垂体腺瘤组织(生长激素腺瘤、泌乳素腺瘤、促性腺激素腺瘤以及裸细胞激素腺瘤各2例)和2例混合的正常垂体组织的基因表达谱.差异表达基因进行筛选和生物信息学分析.芯片结果予以qRT-PCR验证.结果与正常对照组比较,与垂体腺瘤发生关联的基因功能上主要涉及分子结合、凋亡或肿瘤相关、代谢、信号传导、细胞周期、物质运输等多个牛物过程.数个差异基因的表达特异性与亚类腺瘤的发展相关.结论 垂体腺瘤的发生、发展是一多基因、多分子、多通路的网络调控过程.但对于各亚类腺瘤,其分子机制不尽相同.     

利用全基因组芯片初步筛选亚类垂体腺瘤特异性相关基因

目的 探讨人类垂体腺瘤发生、发展中相关基因的表达与功能.方法 应用人全基因组寡核苷酸芯片HG-U133A2.0检测8例垂体腺瘤组织(生长激素腺瘤、泌乳素腺瘤、促性腺激素腺瘤以及裸细胞激素腺瘤各2例)和2例混合的正常垂体组织的基因表达谱.差异表达基因进行筛选和生物信息学分析.芯片结果予以qRT-PCR验证.结果与正常对照组比较,与垂体腺瘤发生关联的基因功能上主要涉及分子结合、凋亡或肿瘤相关、代谢、信号传导、细胞周期、物质运输等多个牛物过程.数个差异基因的表达特异性与亚类腺瘤的发展相关.结论 垂体腺瘤的发生、发展是一多基因、多分子、多通路的网络调控过程.但对于各亚类腺瘤,其分子机制不尽相同.     

利用全基因组芯片初步筛选亚类垂体腺瘤特异性相关基因

目的 探讨人类垂体腺瘤发生、发展中相关基因的表达与功能.方法 应用人全基因组寡核苷酸芯片HG-U133A2.0检测8例垂体腺瘤组织(生长激素腺瘤、泌乳素腺瘤、促性腺激素腺瘤以及裸细胞激素腺瘤各2例)和2例混合的正常垂体组织的基因表达谱.差异表达基因进行筛选和生物信息学分析.芯片结果予以qRT-PCR验证.结果与正常对照组比较,与垂体腺瘤发生关联的基因功能上主要涉及分子结合、凋亡或肿瘤相关、代谢、信号传导、细胞周期、物质运输等多个牛物过程.数个差异基因的表达特异性与亚类腺瘤的发展相关.结论 垂体腺瘤的发生、发展是一多基因、多分子、多通路的网络调控过程.但对于各亚类腺瘤,其分子机制不尽相同.     

LRIG2蛋白表达与人垂体腺瘤侵袭性的研究

目的探索研究LRIG2蛋白表达水平与人垂体腺瘤侵袭性的相关性。方法采用免疫组织化学染色法检测159例垂体标本（8例正常人垂体标本,151例人垂体腺瘤标本）中LRIG2和MMP-9的表达水平,并进行相关性分析。结果LRIG2、在正常人垂体中无阳性反应,LRIG2蛋白表达水平在侵袭性垂体腺瘤比非侵袭性垂体腺瘤显著增高（P〈0.001）,且LRIG2蛋白与MMP-9的表达水平呈正相关（r=0.698,P〈00.001）。结论LRIG2可能作为一个新的垂体腺瘤侵袭性预测指标,检测LRIG2表达水平可指导临床评价垂体腺瘤的侵袭性及制定个体化治疗方案。     

Expression for peroxisome proliferator-activated receptor gamma in pituitary adenomas☆ 38 cases for semi-quantitative immunohistochemical analysis

BACKGROUND： It has been reported that peroxisome proliferator-activated receptor γ （PPAR γ ） is highly expressed in lung cancer, colon cancer, and gastric cancer, as well as other tumors.
OBJECTIVE： To study expression of PPAR γ in pituitary adenomas and analyze the role of PPAR γ in hormonal typing of pituitary adenomas.
DESIGN, TIME AND SETTING： Semi-quantitative immunohistochemistry of pathological specimens. The experiment was conducted at the Department of Neurosurgery, Wuxi Second Hospital Affiliated to Nanjing Medical University between January 2002 and May 2005.
MATERIALS： Surgical resection samples of pituitary adenomas from 38 cases （18 male and 20 female） were analyzed. Eight cases were determined to be invasive pituitary adenomas and 30 cases were non-invasive pituitary adenomas. Hormonal classification of the types of pituitary adenomas revealed somatotrophic adenomas in six cases, corticotrophic adenoma in five cases, prolactinomas in 13 cases, multi-hormone secreting adenomas in six cases, and eight cases of adenoma without altered endocrine function. Five autopsy specimens were collected during the same period from patients of matching age that died from unrelated diseases and were included as normal anterior pituitar3, controls.
METHODS： Cell counts for positive immunohistochemical signals were recorded from histopathological sections. The percentage of positive cells was reported as a semi-quantitative analysis.
MAIN OUTCOME MEASURES： The rate of PPAR γ positive cells in different types of adenoma was based on hormonal levels and invasiveness of pituitary tumor cells.
RESULTS： All tumor biopsies were determined to express PPAR γ. The rate of PPAR γ -positive cells ranged between 8%-65% in the pituitary adenomas. According to hormonal type, PPAR γ expression did not vary between the groups. In addition, there was no significant difference in PPAR γ expression between the non-invasive and invasive pituitary adenomas.
CONCLUSIONS： Human pituitary adenom     

Peroxisome proliferator-activated receptor gamma in the human pituitary gland: expression and splicing pattern in adenomas versus normal pituitary

Pituitary adenomas are slow-growing tumours arising within the pituitary gland. If secreting, they give rise to well-known syndromes such as Cushing's disease or acromegaly; when hormonally inactive, they come to clinical attention often with local mass effects or pituitary deficiency. Peroxisome proliferator-activated receptor gamma (PPARgamma), a nuclear hormone receptor with a key role in fat and glucose metabolism, but also involved in several neoplasia, has recently been detected in pituitary adenomas. In the present study, we evaluated the occurrence and splicing profile of PPARgamma in 43 cases of pituitary adenoma of different subtypes and compared it to 12 normal pituitary glands. By real-time polymerase chain reaction, PPARgamma was expressed as much in adrenocorticotrophic hormone (ACTH)-secreting and ACTH-silent adenomas as in controls, with a moderate underexpression in somatotrophinomas and prolactinomas and overexpression in 54% of nonfunctioning pituitary adenomas (NFPA). There was no apparent qualitative change in the splicing profile of pathological pituitary glands, nor was the presence of specific isoforms with dominant negative effects against PPARgamma detected. Western blotting revealed similar expression levels in the different subgroups of pituitary adenomas and normal glands. Immunohistochemistry confirmed PPARgamma expression in approximately one-half of analysed samples. The intra- and intergroup differences observed in pituitary adenomas may represent new elements in the process of understanding the different clinical responses of Cushing's and Nelson patients to PPARgamma-ligand treatment. Moreover, the higher level of PPARgamma expression detected in the NFPA subgroup may suggest its possible role as a molecular target in these pituitary adenomas, paving the way for investigations on the effectiveness of treatment with thiazolidinediones in such patients.     

Expression of vascular endothelial growth factor in human brain tumors

Compared to normal brain an increased expression of vascular endothelial growth factor (VEGF) has been reported in many types of brain tumors. However, the numbers of samples analyzed and information about the cellular distribution of VEGF have been limited. Here we used novel monochlonal antibodies against VEGF to analyze, using immunohistochemistry, Western blotting and enzyme-linked immunosorbent assay, its expression in 108 human brain tumors that included astrocytic tumors, meningiomas, pituitary adenomas, primary intracranial germ cell tumors and neuronal tumors. The results showed that 37 of 48 astrocytic tumors (77%) and 15 of 19 meningiomas (79%) were immunoreactive for VEGF, consistent with previous reports. However, in contrast to a previous report that analyzed only VEGF mRNA; all of our 15 pituitary adenomas showed specific immunoreactivity for VEGF. We also extended the studies to previously unanalyzed neoplasms: 13 of 15 primary intracranial germ cell tumors (82%), and 7 of 10 neuronal tumors (70%) were immunoreactive for VEGF. Direct protein analysis by Western blotting confirmed the expression of VEGF in those tumors, and showed differential expression of the isoforms of VEGF protein; a pituitary adenoma expressed both VEGF₁₆₅ and VEGF₁₈₉ proteins, a central neurocytoma expressed only VEGF₁₆₅, while an immature teratoma expressed only VEGF₁₈₉. The data herein show that VEGF is expressed in a wide spectrum of brain tumors and suggest differences among tumor entities in the mechanisms of VEGF up-regulation as well as their employment of distinct VEGF isoforms for neovascularization. Received: 26 January 1998 / Revised, accepted: 15 April 1998     

Molecular profiles of inflammatory myopathies

OBJECTIVE: To describe the use of large-scale gene expression profiles to distinguish broad categories of myopathy and subtypes of inflammatory myopathies (IM) and to provide insight into the pathogenesis of inclusion body myositis (IBM), polymyositis, and dermatomyositis. METHODS: Using Affymetrix GeneChip microarrays, the authors measured the simultaneous expression of approximately 10,000 genes in muscle specimens from 45 patients in four major disease categories (dystrophy, congenital myopathy, inflammatory myopathy, and normal). The authors separately analyzed gene expression in 14 patients limited to the three major subtypes of IM. Bioinformatics techniques were used to classify specimens with similar expression profiles based on global patterns of gene expression and to identify genes with significant differential gene expression compared with normal. RESULTS: Ten of 11 patients with IM, all normals and nemaline myopathies, and 10 of 12 patients with Duchenne muscular dystrophy were correctly classified by this approach. The various subtypes of inflammatory myopathies have distinct gene expression signatures. Specific sets of immune-related genes allow for molecular classification of patients with IBM, polymyositis, and dermatomyositis. Analysis of differential gene expression identifies as relevant to disease pathogenesis previously reported cytokines, major histocompatibility complex class I and II molecules, granzymes, and adhesion molecules, as well as newly identified members of these categories. Increased expression of actin cytoskeleton genes is also identified. CONCLUSIONS: The molecular profiles of muscle tissue in patients with inflammatory myopathies are distinct and represent molecular signatures from which diagnostic insight may follow. Large numbers of differentially expressed genes are rapidly identified.     

垂体泌乳素腺瘤垂体特异转录因子表达的实验研究

目的研究垂体特异转录因子pit-1与PRL水平、肿瘤侵袭性的关系,探讨Pit-1在垂体瘤发病机制中的作用。方法免疫组化和RT—PCR分别测定Pit-1蛋白和pit—1 mRNA的表达,分析Pit-1与肿瘤侵袭性的关系。结果与对照组相比,Pit-1在PRL腺瘤中均有高表达,Pit-1、pit-1 mRNA表达与PRL水平呈正相关并与肿瘤侵袭性相关。结论Pit-1及pit-1 mRNA的表达在垂体PRL腺瘤发病机制中发挥一定作用。     

垂体腺瘤中叶酸受体α的表达及意义

目的探讨叶酸受体α（FRα）在垂体腺瘤中的表达情况及其临床意义。方法收集23例垂体腺瘤的新鲜标本,其中无功能性垂体腺瘤13例,功能性垂体腺瘤10例。采用实时荧光定量PCR及Westernblotting分别检测FRα在垂体腺瘤中的mRNA和蛋白表达量。结果实时荧光定量PCR显示：FRα的mRNA相对表达量在无功能性垂体腺瘤中为64．83±43．77．在功能性垂体腺瘤中为1．07±0．51,两者差异有统计学意义（P〈O．05）。Westernblotting结果显示：FRα蛋白在无功能性垂体腺瘤组中明显高表达,而在功能性垂体腺瘤组中不表达（P〈O．05）。结论FRα在无功能性垂体腺瘤中的表达明显高于功能性垂体腺瘤,这为无功能性垂体腺瘤新的分子靶向诊断和治疗提供一定的基础。     

Signalling Pathway Mediated by CXCR7, an Alternative Chemokine Receptor for Stromal-Cell Derived Factor-1α, in AtT20 Mouse Adrenocorticotrophic Hormone-Secreting Pituitary Adenoma Cells

Stromal cell-derived factor (SDF)-1 and its receptor, CXCR4, have been identified in both neurones and glia of many brain areas. Previous studies have mainly focused on the role of SDF-1 and CXCR4 in modulating the hypothalamic-pituitary axis and their possible involvement in the development of pituitary adenomas. An alternative SDF-1 receptor, CXCR7, has recently been identified, but it has not been studied in the context of pituitary adenomas. The present study aimed to investigate the distribution and function of CXCR7 in pituitary adenomas. The expression of CXCR7, normalised to β-actin, was assessed by tissue microarray analysis of 62 adenomas, including 23 growth hormone (GH)-producing adenomas, 22 nonfunctioning adenomas, seven prolactin (PRL)-producing adenomas, six adrenocorticotrophic hormone-producing adenomas and four thyroid-stimulating hormone-producing adenomas. In vitro functional studies used RNA interference (RNAi) and cDNA microarray analysis to evaluate the CXCR7 signalling pathway in AtT-20 mouse pituitary adenoma cells treated with recombinant mouse SDF-1α and transfected with RNAi against Cxcr7 or control RNAi. In tissue microarray analysis, prominent expression of CXCR7 was observed in GH-producing adenomas and PRL-producing adenomas, and in macroadenomas (P < 0.05). Intracellular signalling via CXCR7 up-regulated Bub1, Cdc29 and Ccnb1, and down-regulated Asns, Gpt, Pycr1, Cars and Dars. The present study demonstrates that the SDF-1α/CXCR7 signalling pathway regulates genes involved in cell cycle control, amino acid metabolism and ligase activity, which comprise targets that are distinct from those of CXCR4.     

CT findings of growth hormone secreting pituitary adenomas

The value of high-resolution computed tomography (CT) in the diagnosis of pituitary adenoma has recently been stressed, especially of the coronal view with contrast enhancement. Analysis of the CT scans of 33 growth hormone (GH) secreting pituitary adenomas was done (11 cases of microadenomas, 7 cases of intrasellar adenomas and 15 cases of macroadenomas with suprasellar extension). In macroadenomas, the density was high in five cases, high with isodense portion in two cases, mixed in four cases, isodense in three cases, and isodense and low dense in one case. Six adenomas showed homogeneous density and nine were heterogeneous. After contrast enhancement, two cases showed marked enhancement, ten cases mild and three cases ring enhancement. Margin of adenoma was smooth in nine cases and irregular in six. Among seven cases of intrasellar adenoma one accompanied primary empty sella. In microadenomas ten of eleven cases had hypodense mass inside the normally enhanced pituitary gland. The margin was ill-defined in seven cases and well-defined in three. Eight cases had pituitary height 7 mm or more. Upper surface of the pituitary gland was convex upward in five cases, flat in four and concave in two. Deviation of pituitary stalk was found in seven cases. Bony changes of sellar floor were recognized in three cases. There was a tendency that serum GH level increased with the increment of the size of adenoma. Serum GH levels in adenomas with ring enhancement were lower than those in the homogeneously enhanced adenomas of similar size. One case with marked enhancement showed the highest GH level among all adenomas of the presented series.(ABSTRACT TRUNCATED AT 250 WORDS)