胆囊癌微环境菌群结构特征和功能学的初步研究

Gallbladder cancer is difficulty in early diagnosis, high degree of malignancy and poor prognosis. The occurrence of gallbladder cancer were confirmed with the long-standing closely relationship between gallbladder mucosa chronic inflammation and bacterial colonization factors, however the specific mechanism is unknown. To study the bliary bacteria community and function and search for "pathogen" may be associated with gallbladder cancer. This provides new ideas and direction for clinical diagnosis and treatment of gallbladder cancer. Roche GS FLX System with advantages of high throughput sequencing technology with the digital signal, high data throughput, high sequencing depth and high accuracy, which makes it possible to research the bliary bacteria community and function. Applicants will be the first research on the origin and the characteristics of bliary bacteria community by Roche GS FLX System high-throughput sequencing technology in gallbladder cancer, and combining the early gallbladder cancer genes using high-throughput sequencing technology, and this is the first time systematically expounds the mutation spectrum, drive key genes and signal pathway in occurrence and development of gallbladder cancer, and further studies on the role of gallbladder cancer specific bacteria in gallbladder cancer gene mutation and its possible mechanism. In addition, We were constructed for the first time in liver gallbladder cancer animal model and separate transfer high metastatic phenotype of gallbladder cancer cell subsets GBC-SD/M3 at home and abroad , and related functional verification will be done in this study.

胆囊癌早期诊断困难，恶性程度高，预后极差。胆囊癌的发生被证实与胆囊粘膜长期存在的慢性炎症、细菌定植等因素关系密切，但具体机制不详。对胆囊微环境内菌群稳态的特征和功能进行研究，寻找与胆囊癌发生可能相关的“致病菌”，可为胆囊癌临床诊治提供新的思路和方向。Roche GS FLX System高通量测序技术以其数字化信号、高数据通量、高测序深度、高准确率等优点，使得胆囊微环境菌群谱的研究成为可能。申请者首次将Roche GS FLX System高通量测序技术应用于胆囊癌特征性的微环境菌群及其来源、特征的研究，结合课题组前期系统阐述的胆囊癌基因突变谱、驱动胆囊癌发生发展的关键基因和信号通路，进一步研究胆囊癌特异性菌群在胆囊癌基因突变中的作用及其可能的机制。 此外，课题组前期在国内外首次构建了胆囊癌肝脏转移动物模型和分离出高转移表型胆囊癌细胞亚群GBC-SD/M3进行相关功能学的验证。

胆囊癌早期诊断困难，恶性程度高，预后极差。胆囊癌的发生被证实与胆囊粘膜长期存在的慢性炎症、细菌定植等因素关系密切，但具体机制不详。本研究中，我们发现胆囊癌患者和慢性结石性胆囊炎患者有着相似的稳定和永久的优势物种，在胆道微生物组成和基因功能上有明显的差异。胃链球菌和肠球菌可能在胆囊癌进展中起潜在作用。此外，在胆囊癌患者和慢性结石性胆囊炎组中，宏基因组的物种谱、共丰度、共排除相关和CAZyme代谢谱也有显著差异。体内动物分析发现，肠球菌粪便可促进胆囊癌的进展。