男性非肌层浸润性膀胱癌患者的尿液菌群特征

Characters of urinary microbiota in male patients with non-muscle invasive bladder cancer

目的探讨男性非肌层浸润性膀胱癌患者的尿液菌群特征。方法采用横断面研究方法，选取2017年3-9月我院确诊为非肌层浸润性膀胱癌的男性患者（肿瘤组）和同期健康男性志愿者（对照组）纳入研究。收集两组的人口学特征资料。两组均采集清洁中段尿标本30-50ml，离心留取沉淀物并进行DNA提取、16SrRNA基因扩增和高通量测序。测序原始数据导入QIIME软件，进行质量过滤、操作分类单位聚类和物种分类注释，基于注释结果进行样品微生物α多样性和B多样性分析，LEfSe在线分析两组间有显著差异的菌属。结果肿瘤组和对照组分别纳人26例和18例，肿瘤组吸烟例数明显多于对照组（21例与7例，P〈0．01），其余人口学特征差异均无统计学意义（P〉0．01）。肿瘤组尿液菌群丰度明显高于对照组（Observedspecies指数125．77±69．64与80．38±46．24，P=0．013；Chaol指数142．82±76．74与90．68±47．62，P=0．012；Ace指数147．92±77．68与88．19±45．38，P=0．005），但两组问群落多样性差异无统计学意义（香农指数2．34±0．7与2．27±0．68；辛普森指数0．23±0．12与0．23±0．18；P均〉0．05）。相似性分析结果显示肿瘤组与对照组尿液菌群问分别聚类成两个不同区域，p多样性差异有统计学意义（P=0．009）。LEISe在线分析结果显示，肿瘤组尿液菌群中红色杆菌纲一红色杆菌目．红色杆菌科一红色杆菌属、鞘酯杆菌纲．鞘酯杆菌目一鞘酯杆菌科、不动杆菌属、气单胞菌属和芽孢杆菌属的相对丰度明显高于对照组。结论男性非肌层浸润性膀胱癌患者尿液菌群特征明显不同于男性健康人群，尿液菌群特征与非肌层浸润性膀胱癌的预后可能相关。

Objective To characterize the urinary microbial profile of male non-muscle invasive bladder cancer patients compared to healthy controls. Methods Between March, 2017 and September, 2017,mid-stream urine from 26 non-muscle invasive bladder cancer and 18 non-neoplastic controls were collected by the clean method, then centrifuged and processed for 16S rRNA gene sequencing. Sequencing reads were processed for evaluating alpha diversity and beta diversity using QIIME. LEfSe algorithm was performed to identify potential bacterial genera biomarker. Results The smoking cases were more in tumor group than those in control group（21 vs. 7, P 〈 0.01 ）. The urinary microenvironment of bladder cancer was characterized by increased bacterial richness （Observed species index, Chaol index and Ace index, 125.77 ± 69.64 vs. 80.38 ± 46.24 ; 142.82 ± 76.74 vs. 90.68 ± 47.62 ; and 147.92±77.68 vs. 88.19 ± 45.38, all P 〈 0. 05） and by the enrichment of some bacteria/ genera （ e. g. , Acinetobacter and Anaerococcus）. Significant difference in [3 diversity was found between cancer and non-cancer group （ANOSIM, P = 0. 009）. A clear hierarchical clustering of cancer samples was observed, suggesting a common dysbiosis associated to bladder cancer. Conclusions Patients with non-muscle invasive bladder cancer exhibit a different microbial community compared to non-neoplastic controls, suggesting a possible pathophysiologieal correlation between microbiome and bladder cancer. Urinary microbial community may be associated with the prognosis of NMIBC.