肺腺癌与肺鳞癌患者肠道菌群结构分析

Analysis of gut microbiome in patients with lung adenocarcinoma and lung squamous cell carcinoma

探讨肺腺癌和肺鳞癌患者肠道菌群结构的多样性及构成。采用单中心病例对照研究,连续入组2018年9月至2020年10月就诊于浙江大学医学院附属杭州市第一人民医院的肺癌患者共27例,其中男性15例,女性12例,同期招募健康体检者作为对照组(HC)共20例,其中男性9例,女性11例。收集研究对象新鲜粪便样本及相关临床资料,根据病理诊断结果将肺癌患者分为肺腺癌组(AC,纳入19例患者,其中男性8例,女性11例)和肺鳞癌组(SCC,纳入8例患者,其中男性7例,女性1例)。提取粪便样本基因组DNA,对16S rDNA V3-V4区域进行PCR扩增,通过Illumina MiSeq高通量测序平台进行测序,应用QIIME分析肠道菌群结构。采用方差分析、χ^(2)检验、K-W检验等方法分析3组人群间性别、年龄、α多样性、菌群相对丰度差异等的差异。AC、SCC和HC三组的年龄分别为(58.74±9.27)岁、(63.38±6.12)岁和(55.65±7.79)岁;AC、SCC和HC三组间在性别、年龄上差异无统计学意义(性别和年龄分别为:χ^(2)=5.155,P=0.076;F=2.598,P=0.086)。三组间肠道菌群α多样性无显著性差异(Chao指数和Shannon指数分别为F=0.616,P=0.545;F=2.484,P=0.095),β多样性分析显示AC、SCC与HC三组间肠道菌群结构存在显著性差异(P=0.001)。物种差异分析显示,AC与SCC组均存在优势菌属,巨球型菌属(H=7.855,P=0.020)和Erysipelatoclostridium菌属(H=7.426,P=0.024)在AC患者肠道中富集,而肠球菌属(H=8.400,P=0.015)、韦荣球菌属(H=9.957,P=0.007)和Eubacterium_eligens_group菌属(H=10.514,P=0.005)在SCC患者肠道中富集。肺癌患者存在明显的肠道菌群失调,肺腺癌与肺鳞癌患者肠道菌群结构无显著性差异,但肺腺癌与肺鳞癌具有各自独特的菌群。这种肠道微环境的失衡,对研究不同病理类型肺癌的发生发展具有重要意义。

To investigate the diversity and composition of gut microbiota in patients with lung adenocarcinoma and lung squamous cell carcinoma.A single-center and case-control study was conducted to consecutively enroll a total of 27 lung cancer patients,including 15 males and 12 females,who were seen at the Affiliated Hangzhou First People′s Hospital,Zhejiang University School of Medicine between September 2018 to October 2020.A total of 20 cases of healthy healthy physical examiners,including 9 males and 11 females were recruited as healthy control group(HC)during the same period.Clinical data and stool samples were collected from each participants,and lung cancer patients were divided into lung adenocarcinoma group(AC,19 patients,8 males and 11 females)and lung squamous cell carcinoma group(SCC,8 patients,7 males and 1 females)according to the pathology type.Genomic DNA were extracted to amplify 16S rDNA V3-V4 region,then the Illumina MiSeq high-throughput sequencing platform and QIIME software were used for sequencing and analyzing the structure of the gut microbiota,respectively.Analysis of variance,χ^(2) test,K-W test were used to analyze the differences in age,gender,αdiversity,and relative abundance of microbiota among the three groups.AC,SCC,and HC were aged(58.74±9.27),(63.38±6.12),and(55.65±7.79)years old,respectively.There were no difference in gender and age among the three groups(gender and age are respectively:χ^(2)=5.155,P=0.076;F=2.598,P=0.086).And no significant difference in alpha diversity were found among the three groups(Chao and Shannon index were respectively:F=0.616,P=0.545;F=2.484,P=0.095),whileβ-diversity analysis indicated significant differences in the structure of intestinal flora among AC,SCC and HC(P=0.001).LEfSe analysis showed that AC and SCC both have dominant bacterials.Megasphaera(H=7.855,P=0.020)and Erysipelatoclostridium(H=7.426,P=0.024)were enriched in patients with AC,while Enterococcus(H=8.400,P=0.015),Veillonella(H=9.957,P=0.007),and Eubacterium_eligens_group(H=10.514,P=0.005)were enriched in patients with SCC.Lung cancer patients have gut microbiota imbalance,while lung adenocarcinoma and lung squamous cell carcinoma patients have no significant difference in gut microbiota diversity,but lung adenocarcinoma and lung squamous cell carcinoma have their own unique microbiota.This imbalance of the intestinal microenvironment is of great significance for studying the occurrence and development of different pathological types of lung cancer.