基于高通量测序技术的膀胱癌转录组分子标记的初步筛选

项目名称： 基于高通量测序技术的膀胱癌转录组分子标记的初步筛选

项目编号： No.81460450

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 张淑芳

作者单位： 海口市人民医院

项目金额： 47万元

中文摘要： 膀胱癌是常见的恶性肿瘤之一，其发生发展与基因的变异存在着密切的关系，但还有很多与膀胱癌相关的基因变异未被揭示。 申请人前期对单个膀胱癌患者正常和癌组织的转录组进行测序发现：有 1800 多个基因表达差异显著；300 多个基因的剪接异构体的表达比例发生显著变化，大部分剪接改变的基因之前未见报道，并发现3个耐药基因，其中一个未见报道。 本项目拟采用高通量测序技术，对另外12 例膀胱癌患者的正常和癌组织样品进行转录组测序和生物信息学分析，然后结合 13 例病人的分析结果，筛选出肿瘤特异的新的差异表达基因、肿瘤特异的差异可变剪接基因，基因融合和体突变，并通过大量膀胱癌样品（n>200），采用Real-time PCR方法和Sanger测序技术验证生物信息学分析的结果筛选出候选靶点，并设计细胞及动物水平进行体内外功能实验进行验证，推动膀胱癌分子诊断、肿瘤监测和个性化治疗的发展。

中文关键词： C13_膀胱肿瘤；转录组测序；生物信息学分析；基因变异；分子标记

英文摘要： Bladder cancer is one of the most common malignant tumors, and there is close connection between its development and gene mutation, but many genetic variations associated with bladder cancer have not been revealed. We performed high-throughput transcriptome sequencing on muscle-invasive UCB tissue and found: there were more than 1800 cancer-specific differentially expressed genes between the tumor and none-tumor tissue. Our data also revealed that more than 300 genes showed differential splicing patterns between them, and most of the gene splicing changes had not been reported before, and also three drug-resistance genes were identified. One of them had never been reported in bladder cancer. This project intends to adopt high-throughput sequencing technology to perform transcriptome sequencing and bioinformatics analysis on normal and cancerous tissue samples in another 12 patients with bladder cancer, combining the analysis results of the 13 patients, screens and identifies the new differentially expressed genes , alternative splicing, gene fusion and somatic mutation specific to bladder cancer. At last we would verify our hypothesis and identify the candidate targets by performing reverse transcription-polymerase chain reaction and Sanger sequencing on more than 200 cases of bladder cancer. And furtherly we will design in vivo and in vitro functional experiments to verify them. Our job will surely improve the molecular diagnostics, monitoring and personalized therapy for bladder cancer.

英文关键词： bladder cancer;transcriptome sequencing;bioinformatics analysis;genetic variation;molecular markers