尿路上皮膀胱癌患者泌尿道菌群改变及其对患者预后的影响

Profiling of urinary tract microbiome and its impact on clinical outcome in patients with urothelial bladder cancer

目的研究尿路上皮膀胱癌患者尿液中菌群的变化特征及其对膀胱癌患者预后的影响,为该类患者的治疗提供参考。方法选取2018年11月至2021年11月于我院接受治疗的113例尿路上皮膀胱癌患者作为研究对象,经筛选后最终纳入患者89例。根据WHO 2016年分级标准将患者分为低度恶性潜能乳头状尿路上皮肿瘤组(A组,43例)、低级别乳头状尿路上皮癌组(B组,26例)和高级别乳头状尿路上皮癌组(C组,20例),另外选取同期我院健康体检者50例为对照组,收集所有对象的一般资料,同时收集4组对象中段尿液样本,采用16S rRNA基因高通量测序法检测菌群构成,分析尿液菌群变化特征,并且利用有序变量分析菌群改变与膀胱癌病理分级之间的相关性。结果3组膀胱癌患者性别、年龄、平均身高、平均BMI,吸烟史、酗酒史、高血压史、糖尿病史、冠心病史情况比较差异均无统计学意义(均P>0.05)。和对照组相比,其他3组患者尿液菌群Simpson指数、Shannon指数均显著增加(均P<0.05),其中C组患者Simpson指数高于A组和B组,Shannon指数3组之间差异均有统计学意义(均P<0.05)。和对照组相比,膀胱癌患者尿液中拟杆菌门和放线菌门的丰度无显著变化,但厚壁菌门丰度显著增加,尤其是C组患者达(33.04±5.03)%;膀胱癌患者尿液中变形菌门丰度降低,且随着肿瘤恶性程度的增高而降低(均P<0.05);膀胱癌患者尿液中不动杆菌属丰度无显著变化(P>0.05),但乳杆菌属,假单胞菌属和双歧杆菌属丰度显著降低,且随着恶性程度的增高而降低,尤其是C组患者(均P<0.05)。有序变量回归分析显示,Simpson指数、Shannon指数是膀胱癌预后不良的危险因素,变形菌门、乳杆菌属、双歧杆菌属是保护因素(均P<0.05)。结论膀胱癌患者尿液菌群多样性和构成情况与健康人群有显著区别,菌群多样性、厚壁菌门和变形菌门的改变以及乳杆菌属和双歧杆菌属的减少可能与膀胱癌患者的预后相关。

Objective To observe the change of characteristics of microbiome in the urine of patients with urothelial bladder cancer(BC)and its impact on the clinical prognosis of patients with BC,so as to provide reference for the treatment of such patients.Methods A total of 113 patients diagnosed with urothelial BC who were treated in our hospital from November 2018 to November 2021 were selected,among whom 89 were enrolled according to inclusion/exclusion criteria.According to WHO 2016 standard,the patients were divided into low malignant potential papillary urothelial tumors group(group A,43 cases),low-grade papillary urothelial carcinoma(group B,26 cases)and high-grade papillary urothelial carcinoma(group C,20 cases),with 50 healthy physical examinees in our hospital during the same period as the control group,and the general information of all subjects was collected.The urine samples were collected for detecting the composition of flora using 16S rRNA gene high-throughput sequencing;the characteristics floral changes and the relationship between microbiome changes and BC were analyzed by using ordered variables.Results There were no significant differences in sex,age,average height,average BMI,and histories of smoking,alcoholism,hypertension,diabetes and coronary heart disease among the three groups of BC patients(all P>0.05).Compared with control group,the Simpson index and Shannon index of bacterial flora in the three groups of patients significantly increased(all P<0.05);the Simpson index in group C were higher than in group A and gourp B;the Shannon indexes were different among the 3 groups.Compared with control group,the abundances of Bacteroides and Actinomycetes in patients with BC did not change significantly(all P>0.05),while that of Firmicutes increased significantly,especially in the group C,which was(33.04±5.03)%;the abundance of Proteobacteria decreased in BC patients as the degree of malignancy increased.Compared with control group,there was no significant change in the abundance of Acinetobacter in patients with BC(P>0.05),but those of Lactobacillus,Pseudomonas and Bifidobacterium significantly decreased,especially in group C,along with with the increase of malignancy(all P<0.05).Ordered variable regression analysis showed that the Simpson index,Shannon index were risk factors for poor prognosis of BC,while Proteobacteria,Lactobacillus and Bifidobacterium were protective factors(all P<0.05).Conclusion The diversity and composition of urinary microbiota in patients with BC are significantly different from those in healthy individuals.Clinical outcomes of BC patients may be related to the floral diversity,changes of Firmicutes and Proteobacteria,and decrease of Lactobacillus and Bifidobacterium.