基于高通量测序技术的膀胱癌尿路“核心菌群”特征解析及辅助诊断价值研究

随着高通量测序技术在微生物领域的应用，已有多项研究证实膀胱及尿液中存在多种细菌，从而有学者提出膀胱尿路菌群这一崭新概念。膀胱癌发生发展与尿路菌群结构变化的关系及发挥主要作用“核心菌群”的组成目前尚未阐明。课题组前期对16例膀胱癌患者的尿液、癌及癌旁组织进行了16S rDNA扩增子测序，发现膀胱癌尿液中的菌群结构与健康人存在差异，癌与癌旁组织中菌群结构差异显著，不同病理阶段的尿路菌群特征也不同，对一些可能作为标志物的细菌进行了验证。本课题拟继续增加样本量，完善高通量测序数据，以期明确可能发挥主要作用的“核心菌群”的组成特征，并利用qPCR进行验证；随后选取“核心菌群”中2～4种可培养细菌，分析其及代谢产物对膀胱癌发生发展的关键基因 FGFR3、 TP53突变的影响；最后将通过qPCR分析健康人群和膀胱癌患者尿液中“核心菌群”的分布规律，评估其作为“标志性细菌”用于膀胱癌辅助诊断的价值。

As the application of high-throughput sequencing in microbiology is increasing in popularity, it has been proved that several kinds of bacterial colonies exist in urinary bladder and urine, which is called urinary microbiota. The core urinary microbiota and the association between bladder cancer and urinary microbiota structure changes have not been clarified yet. Previously, we have sequenced the 16S rDNA extracted from urine, bladder cancer tissues and paraneoplastic tissues. The results suggest that there is significant difference of bacterial diversity between the two tissues, and the urinary microbiota characteristics change in different pathologic stages. In our study, we will identify the major composition of the core urinary microbiota by the high-throughput sequencing technology and qPCR analysis. Then we will detect the influence of metabolites of urinary microbiota to FGFR3 and TP53 mutation. Lastly, we will explore the distributing characteristies of core urinary microbiota in the urine from healthy controls, papillary epithelioma patients and bladder cancer patients. We aim to evaluate the value of core urinary microbiota as biomarkers in bladder cancer diagnosis and to investigate the optimal panel of urinary combined detection in bladder cancer.