1. 上海中医药大学附属岳阳中西医结合医院肿瘤科,上海,200437
2. 复旦大学附属肿瘤医院中西医结合科,上海,200032
3. 复旦大学上海医学院肿瘤学系,上海,200032
4. 上海中医药大学基础医学院,上海,201203
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谢国群, 张院辉, 郭晓冬, 等. 基于高通量测序技术的胰腺癌湿热证患者肠道菌群结构差异与功能变异研究[J]. 上海中医药杂志, 2019,53(12):9-16.
XIE Guoqun, ZHANG Yuanhui, GUO Xiaodong, et al. Study on diversity of gut microbiome in pancreatic cancer patients with damp-heat syndrome based on high-throughput sequencing technology[J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(12):9-16.
谢国群, 张院辉, 郭晓冬, 等. 基于高通量测序技术的胰腺癌湿热证患者肠道菌群结构差异与功能变异研究[J]. 上海中医药杂志, 2019,53(12):9-16. DOI: 10.16305/j.1007-1334.2019.12.003.
XIE Guoqun, ZHANG Yuanhui, GUO Xiaodong, et al. Study on diversity of gut microbiome in pancreatic cancer patients with damp-heat syndrome based on high-throughput sequencing technology[J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(12):9-16. DOI: 10.16305/j.1007-1334.2019.12.003.
目的:研究基于高通量测序技术的胰腺癌湿热证患者肠道菌群结构与功能的变化情况,探讨肠道菌群与胰腺癌发病的关联,以期为临床诊治提供新思路。 方法:将2018年1月至2018年12月期间复旦大学附属肿瘤医院和上海中医药大学附属岳阳中西医结合医院收治的31例胰腺癌湿热证患者作为观察组,另选择同期接受健康体检的8例正常人员作为对照组。采集两组人员的粪便样本,提取DNA,建库,经Illumina HiSeq PE150平台实施高通量测序,获得样本的生物学信息,对组间样本进行物种注释分析比较及功能注释分析比较。 结果:①经去冗余后,得到328 787条线粒体开放阅读框(ORFs);两组共有基因为137 849个,观察组独有基因为114 439个,对照组独有基因为60 473个。②在门水平及属水平上,两组间的物种丰度均存在明显差异。在门水平上,观察组菌群丰度中变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、疣微菌门(Verrucomicrobia)、互养菌门(Synergistetes)、纤维杆菌门(Fibrobacteres)的丰度高于对照组,而拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、螺旋体门(Spirochaetes)、念珠菌门(Candidatus)、衣原体门(Chlamydiae)的丰度低于对照组。在属水平上,观察组菌群丰度中艾克曼菌(Akkermansia)、柯林斯氏菌(Collinsella)、链球菌(Streptococcus)、沃氏嗜胆菌(Bilophila)的丰度高于对照组,而双歧杆菌(Bifidobacterium)、普氏菌(Prevotella)、拟杆菌(Bacteroides)、粪杆菌(Faecalibacterium)、梭状杆菌(Clostridium)、罗斯氏菌(Roseburia)的丰度低于对照组。③基于距离矩阵的主坐标分析(PCoA)显示,两组样本的菌群组成结构差异明显。④KEGG数据库功能注释及分析结果显示,观察组在新陈代谢、环境信息处理、细胞加工相关通路上的基因表达水平高于对照组;观察组与对照组的功能丰度分布可以明显区分开。 结论:相较于正常人,胰腺癌湿热证患者体内存在肠道菌群结构失调及功能变异,故监测肠道菌群变化可为临床防治胰腺癌提供有价值的信息。
Objective:To study the diversity of gut microbiome in pancreatic cancer patients with damp-heat syndrome based on high-throughput sequencing technology. This study aims to exploring the role of gut microbiome in the pathogenesis of pancreatic cancer,and providing new ideas for clinical diagnosis and treatment. MethodsThirty-one cases of pancreatic cancer patients with damp-heat syndrome who were treated in our hospital from January 2018 to December 2018 were served as observation group. Another 8 cases of normal people who received a physical examination at the same time were served as control group. Stool samples were collected from two groups of people. DNAs were extracted and the database was constructed. The high-throughput sequencing was performed on the Illumina Hiseq PE150 platform,and then the biological information of the samples was obtained. The species annotation and functional annotation of the samples between the two groups were analyzed and compared. Results:① After deredundancy, 328 787 mitochondrial open reading frames (ORFs) were obtained. The total number of genes in the two groups was 137 849, the unique gene of the observation group was 114 439, and the unique gene in the control group was 60 473.② There was a significant difference in the species abundance between the two groups at the level of the door and the level of the genus. On the level of the door, the abundance of the flora in the observation group was observed, such as the abundance of Proteobacteria, Acinetobacteria, Verrucomicrobia, Synergistertes and Fibrobacteres, which was higher than that of the control group. But the abundance of Bacteroidetes, Firmicutes, Spirochaetes, Candidatus and Chlamydiae was lower than that of the control group. At the genus level, the abundance of Akkermansia, Collinsella, Streptococcus, Bilophila was higher than that of the control group, while the abundance of Bifidobacterium, Prevotella, Bacteroides, Faecalibacterium, Clostridium, Roseburia was lower than that of the control group.③ The principal coordinate analysis (PCoA) based on distance matrix showed that there was significant difference in the composition and structure of flora between the two groups.④ The results of functional interpretation and analysis of KEGG database showed that the gene expression level in metabolism, environmental information processing and cell processing related pathways in the observation group was higher than that in the control group, and the distribution of functional abundance between the observation group and the control group could be distinguished obviously. Conclusion:Compared with normal people,the intestinal flora of pancreatic cancer patients with damp-heat syndrome has obvious differences in diversity,flora structure and function,so monitoring the changes of intestinal flora can provide valuable information for clinical prevention and treatment of pancreatic cancer.
高通量测序技术胰腺癌湿热证肠道菌群组成结构多样性基因功能
high-throughput sequencing technologypancreatic cancerdamp-heat syndromegut microbiomecomposition and structurediversitygene function
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