RTEL1 在脑胶质瘤细胞分化中的作用及机制研究

Nowadays, more attention has been paid to the mechanism of glioma occurrence, and finding the definite therapeutic targets. Our recent work tends to show RTEL 1 gene could be related the glioma occurrence. Additionally, high-levels of RTEL1 expression in glioma tissues is reverse related to gliomas differentiation grades, and related to the expression of mutated p53. However,exactly how RTEL 1 works in order to influence on gliomas differentiation remains unclear, there is a need to acquire further knowledge on it. To elucidate the truth, a series of experiments as follows would be carried out. Firstly, the relation between RTEL 1 expression and gliomas differentiation, and biological behaviors shall be explored, in the experiments where the methods of immunochemistry (ICH) and Western blot assay would be applied. Secondly, at the various stages of glioma cells differentiation, the expression and the transcriptional levels of RTEL 1, and the influences which RTEL 1 expression imposes on glioma cells differentiation would be checked in the experiments where the exons of RTEL 1 could be sequenced to find out all the structure variants at the same time. Furthermore, gene ontology and pathway analysis were used. Thirdly, the correlation between RTEL 1 and p53 is to be discussed. Finally, antibody chips for function classification and gene chips are to be used to detect the correlated biomarks. And the mentioned works are expected to shed light on differentiation therapy,and help explain exactly how RTEL 1 influence on glioma cells differentiation.

探索脑胶质瘤的发生发展机制、寻找确切的治疗靶点是当前脑胶质瘤研究热点。我们前期进行的脑胶质瘤遗传风险研究表明，RTEL1基因可能参与脑胶质瘤的发生，且已发现它在脑胶质瘤组织中高表达并与组织分化呈负相关，与突变型P53蛋白表达相关。然而，它在脑胶质瘤细胞中对分化的影响及机制尚未阐明。本课题拟首先用免疫印迹等方法研究RTEL1的表达与脑胶质瘤细胞分化及肿瘤生物学行为关系；其次，在脑胶质瘤细胞不同分化阶段，对RTEL1基因的外显子测序，发现外显子内所有结构变异；通过PCR、免疫组化等研究RTEL1在脑胶质瘤细胞不同分化阶段的转录、表达，及其表达的高低对细胞分化影响；对差异基因进行基因本体分析、通路分析；再次，探索RTEL1与p53表达相关性；最后应用功能分类的抗体芯片及基因芯片筛选出与RTEL1相互作用的相关分子。本研究将有助于系统地阐明RTEL1参与的脑胶质瘤分化机制，为分化治疗提供新思路。

探索脑胶质瘤的发生发展机制、寻找确切的治疗靶点是当前脑胶质瘤研究热点。我们前期进行的脑胶质瘤遗传风险研究表明，RTEL1 基因可能参与脑胶质瘤的发生，且已发现它在脑胶质瘤组织中高表达并与组织分化呈负相关。然而，它在脑胶质瘤细胞中对发生发展的影响及机制尚未阐明。本项目围绕“RTEL1 基因对脑胶质瘤发生、发展的作用”开展工作，证明了高表达RTEL1基因与胶质瘤患者的不良预后相关；RTEL1基因在胶质瘤发生发展中发挥促癌作用；RTEL1基因除了对端粒具有一定的保护和延长作用外，还可以通过其它的分子机制促进胶质瘤的发生发展，其重要机制之一，即导致一些基因表达异常，并且能调控转录因子ELK1的磷酸化水平发挥其促癌作用。在回答单独 RTEL1 检测，抑或联合其他分子靶标检测，是否能够实现预判脑胶质瘤治疗疗效、初步判断患者生存状况这个问题的过程中，我们亦做了一些尝试。我们探讨了TERT突变与患者生存信息、综合治疗疗效之间的关联，明确了中国人群胶质瘤TERT启动子突变的发生情况，中国人群胶质母细胞瘤（GBM）TERT启动子突变的发生率与文献报道不尽相同；TERT突变、RTL均是判断中国人群胶质瘤患者预后的独立指标。