染色质重塑因子（ARID1A）突变在膀胱移行细胞癌发生中的作用机制研究

项目名称： 染色质重塑因子（ARID1A）突变在膀胱移行细胞癌发生中的作用机制研究

项目编号： No.81502412

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 黄培德

作者单位： 深圳大学

项目金额： 18万元

中文摘要： 染色质重塑调节异常会影响多种基因的正常表达，从而引起癌症等多种疾病的发生。本申请者在膀胱移行细胞癌临床大样本基因组研究中筛选出一个高频突变基因ARID1A。该基因编码产生染色质重塑复合物的一种亚基，我们发现在膀胱移行细胞癌中该基因的多数突变都是终止突变，临床数据显示ARID1A突变与膀胱移行细胞癌患者预后较差相关，提示了该基因突变在膀胱移行细胞癌的发生发展中扮演重要的角色。 .但是目前ARID1A基因突变在膀胱移行细胞癌的发生发展中的作用机制尚不明确。鉴于此，本研究拟在CRISPR/Cas9 构建ARID1A突变型膀胱移行细胞癌细胞系的基础上，通过PCR sanger测序、免疫组化、细胞功能实验、动物活体成瘤和转录组测序等方法，系统深入地探索ARID1A基因突变与膀胱癌的发生和发展关系及其作用的分子机理。为从该基因的作用机制中找到膀胱癌预后诊断与个性化治疗的新靶点奠定基础。

中文关键词： C13_膀胱肿瘤；肿瘤发生；分子机制；染色体重塑；突变

英文摘要： Chromatin remodeling is a dynamic process which promotes the binding of DNA and related factors during DNA replication, repair, restructuring and transcription. Dysregulation of Chromatin remodeling will affect the expression of multiple genes, leading to the occurrence of a variety of diseases such as cancer. .In our former study, by performing massively parallel sequencing in bladder transitional cell carcinomas, we indentified inactivating mutations in ARID1A gene which encoded a subunit of chromatin remodeling complex. We found that most of the mutations found in this gene were truncated mutations. Moreover, clinical data showed that ARID1A mutations correlated with poor prognosis of patients with bladder transitional cell carcinoma. These finding strongly suggested that the mutations in ARID1A may play an important role in the development of bladder transitional cell carcinoma. However, the mechanism whereby which ARID1A promote the development of bladder transitional cell carcinoma still remains unclear due to the lack of systematic functional study. In view of this, we proposed constructing ARID1A mutations bladder transitional cell carcinoma cell lines by CRISPR/Cas9 method, and perform PCR sanger sequencing, immunohistochemical, in vito cell function experiment, in vivo tumorigenesis experiment and transcriptome sequencing to explore the function and mechanism of ARID1A mutations in bladder transitional cell carcinoma. ARID1A may be developed and used as a novel and important diagnosis and prognostic maker for bladder transitional cell carcinoma.

英文关键词： Bladder tumor;Tumorigenesis;Molecular mechanisms ;chromatin remodeling ;mutation