食管鳞状细胞癌肿瘤组织内菌群检测及特征分析

Characterization of Microbiota in Tumor Tissues of Esophageal Squamous Cell Carcinoma

【目的】分析食管鳞癌患者(ESCC组)与健康对照组(HC组)肿瘤组织菌群多样性,筛选出食管鳞癌患者肿瘤组织中特异性细菌。【方法】收集105例食管鳞癌患者肿瘤组织标本和54例健康对照者的食管组织标本,提取标本总DNA,用PCR方法扩增16S rDNA V4区,后行高通量测序,将数据做OTU聚类及多样性分析筛选食管鳞癌组织内特征菌。最后用定量PCR(Q-PCR)扩增方法验证筛选结果。【结果】多样性分析表明ESCC组肿瘤标本和HC组正常食管组织标本内菌群多样性及组成存在明显差异,物种分析表明ESCC组以梭杆菌属(Fusobacterium)、孪生球菌属(Gemella)、奈瑟菌属(Neisseria)、卟啉单胞菌属(Porphyromonas)、链球菌属(Streptococcus)为主,HC组以梭杆菌属(Fusobacterium)、放线杆菌属(Actinobacillus)、链球菌属(Streptococcus)、普雷沃氏菌属(Prevotella)为主,LEfSe分析表明ESCC组卟啉菌属、梭杆菌属丰度升高,且具有统计学差异(P<0.05)。Q-PCR验证结果表明食管鳞癌组织内牙龈卟啉单胞菌、具核梭杆菌拷贝数明显升高。【结论】食管癌肿瘤组织有其特征性的菌群组成,与健康对照组相比,食管鳞癌肿瘤组织中牙龈卟啉单胞菌、具核梭杆菌拷贝数明显升高,可能是其特征性的细菌。

【Objective】To analyze the difference in esophageal microbiome between the patients with esophageal squa⁃mous cell carcinoma(ESCC group)and the healthy controls(HC group),and to screen out the characteristic bacteria in tumor tissues of patients with esophageal squamous cell carcinoma.【Methods】Esophageal tissues were collected from 105 patients with esophageal squamous cell carcinoma and 54 healthy controls.The total DNA of the samples was extracted,and the 16S rDNA V4 region was amplified by PCR,followed by high-throughput sequencing.The data were subjected to OTU clustering and diversity analysis to screen the characteristic bacteria in the ESCC group.Finally,the screening re⁃sults were to be verified by Q-PCR amplification.【Results】Diversity analysis showed that there were differences in diversi⁃ty and composition between the ESCC group and the HC group.Species analysis showed that the main components of microbiome in the ESCC group were Fusobacterium,Gemella,Neisseria,Porphyromonas and Streptococcus.While in the HC group the main components of microbiome were Fusobacterium,Actinobacillus,Streptococcus and Prevotella.The LEfSe analysis showed that the expression of porphyrin and Fusobacteria in the ESCC group increased and the difference is statis⁃tically significant(P<0.05).The results of Q-PCR showed that the presence of Porphyromonas gingivalis and Fusobacteri⁃um nucleatum in the ESCC group significantly increased.【Conclusions】Esophageal cancer has its characteristic flora com⁃position.Porphyromonas gingivalis and Fusobacterium nucleatum in esophageal squamous cell carcinoma tumor tissue are the characteristic bacteria.