摘要
[背景]食管癌是常见的消化道肿瘤,中国是食管癌高发地区。有研究提示,肠道菌群与肿瘤等多种疾病的发生发展相关。受测序读长所限,传统的16S rDNA测序技术只能鉴定到属。[目的]本研究基于PacBio单分子实时(SMRT)测序技术在种水平筛选与食管癌相关的特征性微生物标志。[方法]招募首次诊断为食管癌的新发病例120例,和性别、年龄匹配的健康对照60例。采集所有研究对象的新鲜粪便样本。利用第三代测序PacBio SMRT技术对4例食管癌患者及1:1匹配的健康对照者样本进行16S rDNA全长测序,基于测序结果分析其肠道菌群结构差异。采用PICRUSt软件进行功能预测。通过线性判别分析和群落差异分析筛选差异肠道微生物进行大样本人群验证,识别食管癌相关肠道微生物。[结果]基于测序样本,α多样性分析中食管癌组的Ace、Chao1、Simpson Diversity、Shannon Wiener指数均高于健康对照组(P<0.05),β多样性分析显示食管癌组和健康对照组散点簇分离,二者肠道菌群结构存在差异。在门水平上,食管癌组肠道菌群中变形菌门、拟杆菌门、厚壁菌门丰度升高。在属水平上,食管癌组毛螺旋菌属、巴氏杆菌属、罗斯氏菌属和拟杆菌属相对丰度增加。在种水平上,食管癌组相对丰度增加的微生物是肠杆菌E.20、卵形拟杆菌V975、普氏栖粪杆菌等11种,丰度降低的微生物是皮氏罗尔斯通氏菌、肠杆菌未分类、唾液链球菌JIM87773种。PICRUSt功能注释后发现食管癌组与健康组在丙氨酸、天冬氨酸和谷氨酸代谢(map00250)、肽聚糖生物合成(map00550)、叶酸一碳单位库(map00670)、硫胺素代谢(map00730)、氨基酸的生物合成(map01230)通路上存在差异。对验证人群的分析结果显示,与健康对照人群相比,食管癌人群的肠杆菌E.20、马赛拟杆菌的丰度升高,唾液链球菌JIM8777的丰度降低,差异具有统计学意义(P<0.05)。建立受�
[Background]Esophageal cancer is a common gastrointestinal tumor with a high incidence in China.Some studies suggest that intestinal flora is significantly related to the occurrence and development of tumors and other diseases.Traditional 16S rDNA sequencing technology only provides taxonomic resolution at genus level.[Objective]Based on PacBio single molecule real time(SMRT)sequencing technology to identify characteristic microbial biomarkers associated with esophageal cancer at the species level.[Methods]A total of 120 newly diagnosed cases of esophageal cancer were recruited and 60 healthy patients with matched sex and age were recruited as the control group.Fresh stool samples were collected from all subjects.Full-length 16S rDNA sequencing was performed on samples from 4 patients with esophageal cancer and 1:1 matched healthy controls using the third-generation sequencing PacBio SMRT technology,and the structural differences of intestinal flora were analyzed based on the sequencing results.Function prediction was performed by PICRUSt software.Large population samples were validated by screening different gut microbes by linear discriminant analysis and linear discriminant analysis effect size to identify esophageal cancer-associated gut microbes.[Results]Based on sequencing samples,the results ofαdiversity analysis showed that the Ace,Chao1,Simpson Diversity,and Shannon Wiener indices of the esophageal cancer group were higher than those of the healthy control group(P<0.05),and the results ofβdiversity showed that the scattered clusters of the esophageal cancer group and the healthy control group were separated,which meant that there were differences in the structure of intestinal flora between the two groups.It was found at the phylum level that the abundances of Proteobacteria,Bacteroidetes,and Firmicutes in the intestinal flora of the esophageal cancer group were increased.At the genus level,the relative abundances of Spirospira,Pasteurella,Roxella,and Bacteroides in the esophageal cancer group were inc
作者
万萍
沈伟涛
彭珍燕
刘冉
WAN Ping;SHEN Weitao;PENG Zhenyan;LIU Ran(School of Public Health/Key Laboratory of Environment Medicine Engineering,Ministry of Education,Southeast University,Nanjing,Jiangsu 210009,China)
出处
《环境与职业医学》
CAS
CSCD
北大核心
2022年第9期1011-1020,共10页
Journal of Environmental and Occupational Medicine
基金
国家自然科学基金项目(82173479,81872579)。