新基因C8orf84在膀胱癌中的表达、调控和生物学功能研究

项目名称： 新基因C8orf84在膀胱癌中的表达、调控和生物学功能研究

项目编号： No.81472584

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： 李泽松

作者单位： 深圳大学

项目金额： 75万元

中文摘要： 课题组前期通过转录组分析，筛选出一个在膀胱癌组织中表达显著低于癌旁正常组织的开放阅读框C8orf84。荧光定量PCR及免疫组化检测证实C8orf84在膀胱癌组织表达下调。C8orf84在膀胱癌细胞株中无表达或低表达，并与其启动子区甲基化程度呈负相关。恢复C8orf84表达显著抑制膀胱癌细胞增殖。这些结果提示C8orf84与膀胱癌的发生发展有关，可能是个新候选抑癌基因，但尚未见该基因相关报道。本课题拟在前期研究基础上，采用RT-PCR、Western blot、免疫组化、亚硫酸氢盐基因组测序、染色质免疫共沉淀、慢病毒载体、串联亲和纯化质谱和裸鼠模型等技术方法分析C8orf84在膀胱癌中的表达及其与患者病情的相关性，分析其表达缺失或下调的表观遗传机制，研究C8orf84的生物学功能及发挥作用的信号通路，明确C8orf84在膀胱癌发生发展中的作用，为膀胱癌及其他肿瘤的早期辅助诊断和治疗提供线索。

中文关键词： C13_膀胱肿瘤；C8orf84；肿瘤发生；表观遗传；肿瘤抑制基因

英文摘要： Our prior transcriptome analysis revealed that the expression of open reading frame C8orf84 was lower in bladder urothelial cancer tissues compared to that in matched normal bladder tissues, which was confirmed by quantitative real-time PCR (RT-PCR） and immunohistochemical staining (IHC). C8orf84 was low expression levels or no expression in a panel of bladder cancer cell lines, which was in reverse with the methylation status of CpG islands in its promoter regions. Restore expression of C8orf84 can inhibit proliferation of bladder cancer cells. These preliminary data suggested that C8orf84 play an key role in the bladder cancer carcinogenesis. Thus, we hypothesize that C8orf84 functions as a candidate tumor suppressor gene in bladder cancer. But, there is no report on C8orf84 in MEDLINE database. In order to confirm the hypothesize, we will examine C8orf84 expression by RT-PCR, Western blot and IHC, and determinate the association between its expression and clinicopathological parameters in large bladder cancer samples; then we will investigate the epigenetic regulation mechanism of C8orf84 in baldder cancer by bisulfite genomic sequencing and chromatin immunoprecipitation; last we will investigate the biological function and signaling pathway of C8orf84 in bladder cancer by the lentiviral expression system, nude mice model, tandem affinity purification (TAP) combined with liquid chromatography - tandem mass spectrometry (LC-MS/MS). The purpose is to further clarify the roles of C8orf84 in bladder cancer carcinogenesis and its regulation mechanism, and provide new clues to early diagnosis and gene therapy for bladder urothelial carcinoma and other tumors.

英文关键词： Bladder cancer;C8orf84;Carcinogenesis;Epigenetics;Tumor suppressor gene