基于高通量测序技术研究原发性胃癌患者胃肠道菌群特征_凌宝殿.pdf

第 43 卷 第 3 期2023 年 03 月Vol.43 NO.3MAR.2023赣南医学院学报JOURNAL OF GANNAN MEDICAL UNIVERSITY投稿网址：http：/基于高通量测序技术研究原发性胃癌患者胃肠道菌群特征凌宝殿1，谢志军2，张文娟1，王晓玲1，王芳胜1，胡晓梅1，赖姨梅3，吴何莉2，胡龙华4（1.赣南医学院第一附属医院检验科；2.赣南医学院第一附属医院消化内科；3.赣南医学院第一附属医院病理科，江西 赣州 341000；4.南昌大学医学院第二附属医院检验科，江西 南昌 330000）摘 要：目的：探讨原发性胃癌患者胃肠道菌群种类和分布构成，探究原发性胃癌患者的微生态与癌变发生相关性，并寻找可能存在的菌群生物标志物。方法：根据纳入排除标准，分别收集未经治疗的5例初诊胃癌患者（胃癌组）和5例健康对照人群（对照组）胃黏膜和粪便样本，共20个样本，采用16S rDNA基因高通量测序技术检测胃癌组和对照组胃肠道菌群，通过 PICRUSt2 预测菌群功能差异。结果：操作分类单元（Operational taxonomic units OTU）物种多样性分析显示，胃癌组和对照组胃黏膜样本优化序列数量差异无统计学意义，但胃癌组粪便（WAFB）OTU数明显高于对照组。两组间多样性指数Dominance、Shannon、Chao1、Simpson、Pielou_e及Observed_otus比较差异均无统计学意义，但距离矩阵与（主坐标分析）PCoA分析显示两组菌群结构存在显著差异。线性判别效应分析（LefSe）结果显示，在细菌分类的门水平上，与对照组相比，胃癌组黏膜（WANM）古细菌和拟杆菌门丰度降低，而厚壁菌门和弯曲杆菌门丰度增加，胃癌组粪便（WAFB）的厚壁菌门和放线菌门丰度降低，而变形杆菌门、拟杆菌门和弯曲杆菌门丰度增加。在属水平上，胃癌组大肠埃希菌志贺菌属（Escherichia-Shigella）丰度高于对照组，而拟杆菌属（Bacteroides）等益生菌则低于对照组。功能基因预测结果显示，胃癌组与对照组菌群代谢通路存在差异。结论：胃癌患者胃肠道菌群多样性和结构发生了特征性改变，其中在门水平上，无论是胃黏膜菌群，还是肠道菌群，其拟杆菌门和变形杆菌门为具有显著差异的标志性菌群，在属水平上，胃癌患者大肠埃希菌志贺菌属（Escherichia-Shigella）丰度显著增高，而拟杆菌属（Bacteroides）等益生菌则显著降低。大肠埃希菌志贺菌属（Escherichia-Shigella）可能作为本地区胃癌患者潜在的生物标志物。关键词：胃黏膜菌群；肠道菌群；胃癌；16S rDNA中图分类号：R735.2 文献标志码：A 文章编号：1001-5779（2023）03-0228-07 DOI:10.3969/j.issn.1001-5779.2023.03.002开放科学（资源服务）标识码（OSID）：Study on the characteristics of gastrointestinal flora in patients with primary gastric cancer based on high-throughput sequencing technologyLING Bao-dian1，XIE Zhi-jun2，ZHANG Wen-juan1，WANG Xiao-ling1，WANG Fang-sheng1，HU Xiao-mei1，LAI Yi-mei3，WU He-li2，HU Long-hua4（1.Department of Clinical Laboratory，The First Affiliated Hospital of Gannan Medical University；2.Department of Gastroenterology，The First Affiliated Hospital of Gannan Medical University；3.Department of Pathology，The First Affiliated Hospital of Gannan Medical University，Ganzhou，Jiangxi 341000；4.Department of Clinical Laboratory，The Second Affiliated Hospital of Nanchang University Medical College，Nanchang，Jiangxi 330000）Abstract :Objective :To explore the types and distribution composition of gastrointestinal flora in patients with primary gastric cancer，clarify the synergistic rule of gastrointestinal flora，explore the microecological carcinogenic mechanism in patients with primary gastric cancer，and search for potential biomarkers of bacterial flora.Methods :According to the inclusion and exclusion criteria，Gastric mucosa and fecal samples were collected from 5 newly diagnosed gastric cancer patients and 5 healthy control cases without prior treatment，with a total of 20 samples，16S rDNA gene high-throughput 基金项目：江西省卫生计划生育委员会科技计划项目（20195406）作者简介：凌宝殿，男，硕士在读，研究方向：临床微生物耐药性研究。E-mail：通信作者：胡龙华，男，本科，教授，博士生导师，研究方向：临床细菌致病及耐药机制研究。E-mail：L 2283 期凌宝殿，等 基于高通量测序技术研究原发性胃癌患者胃肠道菌群特征投稿网址：http：/sequencing technology was used to analyze the gastrointestinal flora，and PICRUSt2 predicted the functional differences of the flora.Results :Operational taxonomic units（OTU）analysis showed that there was no statistical difference in the number of optimized sequences of gastric mucosa samples between gastric cancer patients and normal control group，but the number of OTU in gastric cancer feces（WAFB）group was significantly higher than that in control group，indicating that the species richness of intestinal flora in gastric cancer group was greater than that in control group.The comparison of Alpha diversity index between the two groups，Shannon，Chao1，Simpson，Pielou_e and Observed_otus showed no statistical significance，but the distance matrix and PCoA analysis showed significant differences in microflora structure between the two groups.Linear discriminant effect analysis（LefSe）results showed that，at the phylum level of bacterial classification，the abundance of archaea and bacteroidetes decreased in gastric carcinoma mucosa（WANM）group，while the abundance of firmicutes and campylobacter increased；the abundance of firmicutes and actinomycetes decreased in gastric carcinoma feces（WAFB）group，while the abundance of proteobacteria，bacteroidetes and campylobacter increased.At the genus level，the abundance of Escherichia-Shigella in gastric cancer patients was significantly higher than that in the control group，while the abundance of Bacteroides and other probiotics was significantly lower than that in the control group.The results of PICRUSt2 functional gene prediction showed that there were differences in the metabolism of bacteria between gastric cancer patients and healthy control group.Conclusion :The diversity and structure of gastrointestinal flora in patients with gastric cancer were characterized by the decrease of the abundance of Bacteroides and the increase of Escherichia-Shigella and other opportunistic pathogens.Escherichia-Shigella may be a potential biomarker for gastric cancer patients in this region.Key words :Gastric mucosal microflora；Gut flora；Gastric cancer；16S rDNA胃癌（Gastric cancer，GC）是全球第五大流行和第四大致命的恶性肿瘤，占所有癌症相关死亡人数的7.7%1。人体胃肠道定植大量微生物，微生物之间互相拮抗互为制约，维持相对稳定的微生物微环境，组成一个复杂的生态系统。机体胃肠道中的微生物在多种生理过程中发挥着重要作用，参与了能量代谢、营养吸收、肠道免疫系统的成熟以及防止病原体感染2，也参与疾病的发生和发展。微生物群的改变可