基于16S rRNA和非靶向代谢组测序分析结直肠癌患者肠道菌群及代谢物的变化

Changes in gut microbiota and metabolites in colorectal cancer patients based on 16S rRNA and non-targeted metabolome sequencing

目的采用16S rRNA和非靶向代谢组测序分析的方法探讨结直肠癌患者肠道菌群和其代谢产物的变化。方法新鲜粪便为实验样品,收集10个结直肠癌患者组样本和6个正常对照组样本进行16S rRNA测序,分析肠道菌群的组成、多样性和物种差异,并对预测差异肠道菌群代谢功能;取8个结直肠癌组样本和6个正常对照组样本进行LC-MS非靶向代谢组学测序,分析代谢产物变化情况并进行功能预测。结果结直肠癌组的菌群多样性较正常对照组无显著差异(P>0.05)。结直肠癌组的优势菌群为疣微菌门,结直肠癌组葡萄球菌、嗜冷杆菌属、毛螺菌科NK4A136、梭状芽孢杆菌vadinBB60和CAG-56相对丰度增加(P<0.05),CAG-352、瘤胃球菌属、粪球菌属和挑剔真杆菌相对丰度降低(P<0.05)。KEGG富集分析发现,差异菌群在环境信息处理、遗传信息处理和新陈代谢等通路中发挥重要作用。非靶向代谢组学测序检测出两组样本有367种代谢物存在显著差异,对差异代谢物进行功能预测发现与咖啡因、嘌呤、氨基糖和核苷酸糖和生物素等的代谢相关。结论结直肠癌患者的肠道菌群和代谢物与健康个体存在差异,结直肠癌与肠道菌群和代谢途径可能存在一定关联。

Objective To observe the changes in intestinal flora and its metabolites of patients with colorectal cancer(CC)by using 16S rRNA and non-targeted metabolome sequencing analysis.Methods Ten CC samples and 6 normal control samples were collected for 16S rRNA sequencing.The composition,diversity and species differences of intestinal flora were analyzed,and the metabolic function of differential intestinal flora was predicted.Eight CC samples and 6 normal control samples were subjected to LC-MS non-targeted metabolomics sequencing to analyze the metabolic changes and predict the function.Results The floral diversity in the CC group was not significantly different from that in the normal control group(P>0.05).In the CC group,the dominant phylum was Verrucomicrobia;the relative abundances of Staphylococcus,Psychrobacter,Lachnospiraceae_NK4A136_group,Clostridia_vadinBB60_group and CAG-56 increased(P<0.05),while those of CAG-352,Ruminococcus,Coprococcus and Eubacterium_eligens_group decreased(P<0.05).KEGG enrichment analysis found that differential microflora plays important roles in pathways such as environmental information processing,genetic information processing and metabolism.Non-targeted metabolomics sequencing detected significant differences in 367 metabolites in the two groups of samples,and the functions of differential metabolites was found to be associated with the metabolisms of caffeine,purine,amino sugar and nucleotide sugar and biotin.Conclusion The intestinal flora and metabolites of CC patients differ from those of healthy individuals,and there may be some association between CC and the intestinal flora and metabolic pathways.