Bacterial community characterization of intestine and its influence on the occurrence and development of colorectal cancer and precancerous lesions in Ningbo population
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摘要: 目的 探讨肠道细菌在健康人类、大肠肿瘤患者和大肠息肉患者之间的差异,分析微生物对大肠息肉及大肠肿瘤的发生、进展的影响。 方法 样本来源于常规做肠镜检查患者的粪便以及术前愿意参加实验的患者,分为3大组:肠镜检查健康组、息肉组和大肠腺癌组,同时以高血压为独立的考察因素,健康组和息肉组又各分为高血压和非高血压2个亚组,样本经过DNA提取后,16SrRNA基因全长扩增和V3、V4区PCR扩增,采用DGGE预评判提取DNA质量,采用高通量测序技术分析各样本中细菌的种类和丰度,并通过主成分分析探讨不同分组患者肠道细菌的多样性和群落结构差异。 结果 不同分组患者的肠道细菌具有较高的多样性,在获得的所有OTUs中,厚壁菌门、拟杆菌门、变形菌门、梭杆菌门分别占44.9%、28.4%、18.6%和5.5%,这4门的OTUs累计占97.4%。这些构成了大多数样本中的优势菌,但不同样本中优势细菌的组成有显著的区别。各分组的多样性指数统计分析发现,不同组之间的多样性指数差异不显著。健康人肠道细菌群落结构与肿瘤患者有着显著差异,结肠息肉患者的肠道细菌群落结构与健康人群的肠道细菌群落结构差异显著。健康组中非高血压和高血压组的肠道细菌群落结构差异无统计学意义,结肠息肉组中非高血压和高血压组的肠道细菌群落结构差异无统计学意义。 结论 肠道菌群的变化可能影响或导致结肠息肉和结肠肿瘤的发生、发展,观察肠道菌群的变化有助于研究结肠癌发展的微环境及机制,而高血压对健康人和肠道息肉患者的肠道菌群变化的无显著影响。Abstract: Objective To compare the different of bacterial community structure in the intestinal ecosystem of normal subjects with patients with colorectal carcinoma or polyps,and explore the effects of gut microbiota on the occurrence and development of colorectal cancer and precancerous lesions. Methods DNA samples were collected from the stool of volunteers undergoing colonoscopy.The participants were assigned into healthy group,polyps group and colorectal adenocarcinoma group according the colonoscopy.Taking high blood pressure as an independent factor,the participants of both healthy group and polyps group were assigned into hypertension subgroups and non-hypertension subgroups.DNA was prepared for PCR amplification of 16S rRNA genes and its V3-V4 regions.Then the DGGE was used to pre-evaluate DNA quality.The species and abundance of bacteria in each sample were analyzed by high-throughput sequencing.The differences of bacterial diversity and community structure of different groups were analyzed by principal component analysis. Results The intestinal bacteria among these groups had higher diversity.Based on the obtained operational taxonomic units(OTUs),Firmicutes(44.9%),Bacteroidetes(28.4%),Proteobacteria(18.6%) and Fusobacteria(5.5%) accounted for 97.4% of total OTUs,and made up the predominant bacteria in most samples,but the composition of the dominant bacteria in different samples was significantly different.The diversity index of each group showed no significant difference among different groups.The bacterial community structure of intestinal bacteria in healthy people was significantly different from that in cancer patients.The intestinal bacterial community structure of colon polyp was significantly different from the intestinal bacterial community structure of healthy people.There was no significant difference in the bacterial community structure between the non-hypertensive and hypertensive groups from the healthy group.There was no significant difference in colonic structure between the non-hypertensive and hypertensive groups. Conclusion The changes of intestinal microflora may affect or lead to the occurrence and development of colonic polyp and colonic tumors.It is helpful to study the microenvironment and mechanism of colonic cancer development,and the effect of hypertension on intestinal microflora of healthy people and intestine has no significant difference.
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Key words:
- Intestinal bacteria /
- Bacterial community structure /
- Tumor /
- Adenoma /
- High-throughput sequencing
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