【作者】 王爱东；

【导师】 缪竞诚；

【作者基本信息】 苏州大学 ， 免疫学， 2003， 硕士

【摘要】 [目的] 建立人脑胶质瘤恶性进展相关基因表达谱，分析其中与细胞凋亡相关基因的表达差异，并选择有进一步研究价值的基因在不同级别胶质瘤和正常脑组织临床标本进行验证，为探讨凋亡相关基因在胶质瘤恶性进展中的作用奠定基础。[方法] (1)标本收集：收集同一患者初发和两次复发的胶质瘤组织，其病理形态学诊断依次为：神经节胶质瘤(WHOⅡ级)、间变性星型细胞瘤(WHOⅢ级)和多形性胶质母细胞瘤(WHOⅣ级)。同时取得了第二次复发时远离肿瘤的正常脑组织作为正常对照。(2)cDNAmicroaray技术检测三份肿瘤组织和正常脑组织中差异基因的表达，建立胶质瘤恶性进展相关基因表达谱。(3)根据表达谱芯片结果结合GeneBank检索资料分析其中凋亡相关基因的表达变化。(4)RT-PCR和Dot blot检测部分细胞凋亡相关基因在不同级别胶质瘤和正常脑组织中的表达。[结果] (1)三份肿瘤组织中至少有一份与正常脑组织相比有2倍差异的基因982个，肿瘤组织与正常脑组织分别比较，差异基因分别有403、451和386个。依此建立了胶质瘤恶性进展相关基因表达谱。(2)细胞凋亡相关基因31个，其中肿瘤组织与正常组织具差异的17个，包括凋亡抑制因子TNFAIP3、BIRC2等上调和凋亡促进因子CRADD、NADE等下调共同参与了胶质瘤恶性进展。(3)RT-PCR结果显示TNFAIP3在胶质瘤组织中表达升高，ENC1表达下降，与芯片结果一致；Dot blot结果表明肿瘤组织中SLI基因表达上调，CDC27、CRADD、SAMAN1、CASP1表达下调。[结论]建立胶质瘤恶性进展相关基因表达谱，对进一步阐明胶质瘤发生、发展的胶质瘤恶性进展相关基因表达谱中凋亡相关基因的初步研究中文摘要分子机制具有重要价值;在肿瘤组织中TNEAIP3、SLI等基因表达上调，ENCI、CDC27、CRADD、SAMANI、CASPI等表达下调，这些细胞凋亡相关基因共同参与了胶质瘤的恶性进展，为胶质瘤的诊断和治疗提供新的思路和靶点。经初步检索，SU和SAMANI在胶质瘤中的表达变化未见报道，值得进一步研究。更多还原

【Abstract】 Objective To establish the gene expression profile associated with the malignantprogression of glioma. Detecting the apoptosis-related genes expression patterns of human glioma specimens and normal brain tissues for helping us to study the molecular mechanisms of malignant progression of glioma.Methods (1) Collecting primary tumor specimens, tumor specimens of two recurrenceand normal brain tissue of an individual patient. The classifications of tumors are ganglioglioma (WHO II), anaplastic astrocytoma (WHO III), and glioblastoma (WHO IV), respectively. (2) Detecting the gene expression patterns of above four tissues by cDNA microarray, and establishing the gene expression profile associated with malignant progression of glioma. (3) Analyzing the apoptosis-related genes based on the results of cDNA microarray and information from GeneBank. (4) Detecting the apoptosis-related genes expression patterns of different grade glioma tissues and normal brain tissues with RT-PCR and Dot blot for verifying the chip results.Results (1) By comparing the signals of normal brain tissue, there were 403,451, and 386differentially expressed genes more than 2-fold of three tumor specimens, respectively, and the total 982 genes showed more than 2-fold difference. (2) There were 31 differentially expressed apoptosis-related genes, and 17 genes comparing three tumor tissues with the normal one. The apoptosis inhibitors as TNFAIP3, BIRC2 and inducing factors as CRADD, NADE were involved in the malignant progression of glioma. (3) Proved by RT-PCR, TNFAIP3 gene was up-regulated in tumor tissues, and ENC1 genes was down-regulated. In Dot blot study, SLI gene was found up-regulated and CDC27 CRADD SAMAN1 CASP1were down-regulated in tumor tissues compared with nomal brain tissues.Conclusions Established gene expression profile associated with malignant progressionof glioma will be important for us to clarify the molecular mechanisms of the development of glioma. The apoptosis-related genes were involved in malignant progression of glioma, which may provide us new strategies and targets for glioma diagnosis and treatment.更多还原