高通量测序分析不同病理类型食管癌患者食管菌群的特征

High-throughput sequencing in analyzing the features of esophagus flora in patients with different pathological types of esophageal cancer

目的探讨不同病理类型食管癌患者的食管菌群特征。方法 37例食管癌患者依据手术组织病理分为鳞癌组29例和非鳞癌(腺癌和神经内分泌癌)组8例,对细菌16SrRNA基因序列V4高变区扩增,用Illumina HiSeq测序,可操作分类单元聚类分析2组菌群结构、丰度和多样性。结果非鳞癌组Chao1指数(514.790±257.529)、Shannon指数(5.929±1.015)和Simpson指数(0.950±0.027)与鳞癌组(532.636±251.516、5.661±0.962、0.934±0.051)比较差异均无统计学意义(P>0.05),2组菌群丰度相似;非度量多维尺度分析显示2组样本分布有交叉,不能完全区分,2组菌群多样性无明显差异。在门水平,非鳞癌组放线菌门、绿菌门、脱铁杆菌门、梭杆菌门丰度较鳞癌组增多(P<0.05),其余菌门2组比较差异无统计学意义(P>0.05)。差异显著的29个属中,鳞癌组短杆菌属、弗兰克氏菌属、纤维菌属、桥杆菌属、香味菌属、糖杆菌属、芽孢八叠球菌属、寡源杆菌属、动胶杆菌属、硫曲菌属、军团菌属、盐单胞菌属缺失(丰度为0),与非鳞癌组比较差异有统计学意义(P<0.05)。结论通过Illumina HiSep高通量测序技术建立本地区食管癌微生态基因集,鳞癌和非鳞癌食管菌群多样性无明显差异,但在门水平和属水平上存在明显差异。

Objective To analyze the characteristics of esophageal flora in patients with different pathological types of esophageal cancer.Methods Thirty-seven esophageal cancer tissue samples were divided into squamous cell carcinoma(SCC)group(n=29)and non-SCC group(n=8).The 16 SrRNA gene sequence V4 hypervariable region of bacteria was amplified and sequenced by Illumina HiSeq.Cluster analysis of operational taxonomic units was used to analyze the structure,abundance and diversity of bacteria in two groups.Results There were no significant differences in the Chao1 index(514.790±257.529 vs.532.636±251.516),Shannon index(5.929±1.015 vs.5.661±0.962)and Simpson index(0.950±0.027 vs.0.934±0.051)between non-SCC group and SCC group(P>0.05),indicating that the abundance was similar between two groups.Non-metric multidimensional scale analysis showed that the sample distribution of two groups was not completely distinguished,indicating a similar diversity of bacteria.At the phylum level,the abundances of Actinobacteria,Chlorobi,Deferribacteres and Fusobacteria were higher in non-SCC group than those in SCC group(P<0.05),and the other two bacteria showed no significant differences between two groups(P>0.05).In 29 genera with great differences,Brevibacterium,Frankella,Cellulobacter,Bridges,Fragrance,Glycobacteria,Bacillus spores,Oligobacteria,Klebsiella,Sulfur Bacteria,Legionella and Halomonas were missing in SCC group(abundance:0),showing significant differences from those in non-SCC group(P<0.05).Conclusion Metagenomic sequencing technology can be used to establish the genome of esophageal cancer.SCC and non-SCC show significant differences in the levels of the phylum and genus but not in the diversity.