MicroRNA调控自噬对胶质母细胞瘤放疗敏感性的作用和机制研究

项目名称： MicroRNA调控自噬对胶质母细胞瘤放疗敏感性的作用和机制研究

项目编号： No.81472346

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 赵鹏

作者单位： 浙江大学

项目金额： 72万元

中文摘要： 自噬近年来被认为是恶性肿瘤放疗抗拒的重要机制之一。我们前期研究证实抑制自噬的发生能增强胶质母细胞瘤的抗肿瘤治疗效果，通过miRNA芯片分析胶质母细胞瘤放疗抗拒细胞和亲本细胞，发现4个可能与自噬相关的候选miRNA表达存在差异，提示miRNA可能是自噬影响胶质母细胞瘤放疗抗拒的关键因子。本研究拟通过实时定量PCR鉴定miRNA表达差异，运用生物信息软件分析这些候选miRNA与自噬通路相关的靶基因并验证，利用miRNA 模拟/抑制、过表达/沉默靶基因的方法观察胶质母细胞瘤放疗敏感性和自噬变化情况，并通过皮下移植瘤和原位瘤动物模型进一步确认。最后，分析临床标本的候选miRNA与相应的靶基因、自噬相关基因表达的相关性，以及与放疗疗效、临床分期和预后的关系。本项目能阐明自噬影响胶质母细胞瘤放疗敏感性的具体分子机制，为肿瘤治疗抗拒提供新的治疗靶点，具有重要的临床意义。

中文关键词： 胶质母细胞瘤；自噬；微小RNA；放射抗拒

英文摘要： Autophagy is considered to be one of the important mechanisms of resistance to radiotherapy. Our previous studies have demonstrated inhibition of autophagy can enhance anti-tumor treatment in glioblastoma. miRNA microarray analysis revealed that there are levels of different miRNA between radioresistant glioblastoma cells and the parental cell from which they were derived, suggesting that miRNA may be the key factor that autophagy affect radiosensitivity. In this study, quantitative real time PCR was used to evaluate the expression the different miRNA. Using integrated bioinformatic alogorithms to predict the potential target gene associated with autophagy of the miRNAs. We transfected glioblastoma cells with miRNA mimics or inhibitor, overexpression target gene or siRNA plasmid and evaluated their effect on cell radiosensitivity and autophagy in vitro and in vivo (subcutaneous and in situ tumor xenograft). Finally, we investigate the correlation among the candidate miRNA, target gene and autophagy activity in human glioblastoma tissues, and its relationship with radiotherapy effect, clinical staging and prognosis. This study is important to elucidate the molecular mechanism of autophagy affect radiosensitivity in glioblastoma and to provide the novel target of cancer treatment, which is meanfingful in clinical.

英文关键词： glioblastoma;autophagy;miRNA;radioresistance