高通量测序分析乳腺癌微生物群落特征

High-throughput sequencing analysis of breast cancer microbial community characteristics

目的分析乳腺癌患者肿瘤组织与癌旁组织微生物菌群多样性,筛选乳腺癌患者肿瘤组织中的特异性细菌。方法分析2021年2月至9月河南省肿瘤医院行乳腺癌根治术切除的20例乳腺癌患者的肿瘤组织(TT组)与配对瘤旁组织(TN组),采用Illumina HiSeq高通量测序技术对乳腺癌患者组织样本进行细菌V3-V4区的16S rDNA测序及生物信息学分析,采用t检验进行计数资料比较。结果在门水平上,TT组与TN组的微生物菌群主要由变形菌门、放线菌门、厚壁菌门和拟杆菌门组成,累计含量>60%。微生物多样性分析显示,Chao1指数TN组比TT组为10167.59±2929.38比9044.89±2735.11(t=1.09,P>0.05),Ace指数TN组比TT组为12571.96±2859.65比11704.53±2855.75(t=0.81,P>0.05)。Beta多样性显示两组菌群结构差异有统计学意义(R=0.27,P<0.001)。门水平差异分析显示,变形菌门(TN组比TT组为42.63±16.76比52.08±9.53,t=-0.65)、放线菌门(TN组比TT组为6.13±2.55比9.02±1.77,t=-1.03)等在癌组织中明显增加(P<0.05),厚壁菌门(TN组比TT组为10.62±9.40比4.40±0.95,t=3.41)等明显减少(P<0.05)。线性判别分析(LDA)显示TT组中α变形菌纲、根瘤菌目、和变形菌门等为显著性差异菌,而TN组中链球菌属和假单胞菌属等为显著性差异菌。结论乳腺癌患者癌组织与癌旁组织的微生物群落结构存在明显差异,其中假单胞菌属、链球菌属等为差异微生物有助于筛选标志物。

Objective To analyze the microbial community characteristics in tumor tissues and adjacent nontumor tissues of breast cancer patients,and screen specific bacteria in tumor tissues.Methods Tumor tissues(TT group)and adjacent nontumor tissues(TN group)from 20 breast cancer patients in Henan Cancer Hospital were collected.Illumina HiSeq high-throughput sequencing technology was used to conduct 16S rDNA sequencing of the bacterial V3-V4 region and bioinformatics analysis.T-tests was used to analyze the data.Results At the phylum level,the microbial community mainly consists of Proteobacteria,Actinobacteria,Firmicutes,and Bacteroidetes,accumulated content>60%.There is no statistically significant difference in bacterial abundance,characterized by the Chao1 index TN vs.TT(10167.59±2929.38 vs.9044.89±2735.11,t=1.09)and Ace index TN vs.TT(12571.96±2859.65 vs.11704.53±2855.75,t=0.81),between breast cancer tissue and adjacent tissue(P>0.05).The beta diversity analysis demonstrated a statistically significant difference in microbial community structure between TT group and TN group(R=0.27,P<0.001).The phylum-level difference analysis showed a significant increase in Proteobacteria(TN vs.TT:42.63±16.76 vs.52.08±9.53,t=-0.65),Actinobacteria(TN vs.TT:6.13±2.55 vs.9.02±1.77,t=-1.03)and other phyla in cancer tissues(P<0.05),while a significant decrease was observed in Firmicutes(TN vs.TT:10.62±9.40 vs.4.40±0.95,t=3.41)in cancer tissues(P<0.05).Linear discriminant analysis(LDA)revealed that Alphaproteobacteria,Rhizobiales,Proteobacteria,etc.in the TT group were identified as differential bacteria.Streptococcus and Pseudomonas,etc.were identified as differential bacteria in the TN group.Conclusion There is a significant difference in the microbial community structure between cancer tissues and adjacent nontumor tissues in breast cancer patients.Pseudomonas,Streptococcus,etc.are differential microorganisms that are helpful for screening markers.