高通量测序技术分析化疗对食管癌患者肠道菌群的影响及其与化疗效果的关系

Analysis of the effect of chemotherapy on intestinal flora of patients with esophageal cancer by high-throughput sequencing technology and the relationship of intestinal flora with the efficacy of chemotherapy

目的探讨化疗对食管癌患者肠道菌群的影响及其与化疗效果的关系。方法选取2021年11月至2022年3月河南省南阳市中心医院收治的食管癌患者12例。收集其化疗前及化疗2个周期后的粪便样本,采用16S rRNA高通量测序技术分析粪便样本中的微生物多样性和组成差异,以及不同化疗效果组之间的菌群差异。结果在门水平上,化疗前后食管癌的肠道菌群均主要由厚壁菌门、放线菌门、拟杆菌门和变形菌门组成,占总群落的96.0%以上。化疗后粪便样本中表征肠道菌群丰富度的Chao1指数和Observed-species指数以及表征肠道菌群多样性的Shannon指数和Simpson指数均明显高于化疗前样本[(1916±1260)比(1005±302)、(1672±1073)比(869±279)、(7.0±1.3)比(5.9±0.7)、(0.96±0.03)比(0.93±0.06)],差异均有统计学意义(均P<0.05)。Beta多样性分析结果显示化疗后与化疗前的粪便样本中的菌群结构差异有统计学意义,组间差异大于组内差异(R=0.089,P=0.049)。线性判别分析效应大小分析显示,肠道菌群厚壁菌门、梭杆菌科等在临床获益组患者中明显减少(P<0.05),乳酸杆菌属、罗氏菌属在临床获益组患者中则明显增加(P<0.05)。结论食管癌患者化疗前后肠道微生物群落结构存在明显差异,临床获益患者的肠道微生物群中以罗氏菌属、乳酸杆菌为主的产短链脂肪酸菌属明显增多。

Objective To explore the effect of chemotherapy on intestinal flora of patients with esophageal cancer by high-throughput sequencing technology and the relationship of intestinal flora with the efficacy of chemotherapy.Methods Totally 12 patients with esophageal cancer in Nanyang Central Hospital,Henan Province from November 2021 to March 2022 were enrolled,and their fecal samples before and after 2 cycles of chemotherapy were collected.The 16S rRNA high-throughput sequencing technology was used to analyze the diversity and composition of microorganisms in fecal samples,as well as the differences of flora between groups with different chemotherapy effects.Results At the phylum level,the intestinal flora of esophageal cancer was mainly composed of Firmicutes,Actinobacteria,Bacteroidetes and Proteobacteria before and after chemotherapy,accounting for more than 96.0%of the total community.The Chao1 index and Observed-species index representing the abundance of intestinal flora and the Shannon index and Simpson index representing the diversity of intestinal flora of the samples after chemotherapy were significantly higher than those of the samples before chemotherapy[(1916±1260)vs(1005±302),(1672±1073)vs(869±279),(7.0±1.3)vs(5.9±0.7),(0.96±0.03)vs(0.93±0.06)](all P<0.05).Beta diversity analysis showed that there was a statistically significant difference in the flora structure of fecal samples before and after chemotherapy,and the difference between groups was greater than that within groups(R=0.089,P=0.049).The effect size analysis of linear discriminant analysis showed that intestinal flora Firmicutes,Fusobacteriaceae and others significantly reduced in the patients of the clinical benefit group(P<0.05),while Lactobacillus and Rochella significantly increased in the patients of the clinical benefit group(P<0.05).Conclusions There are significant differences in the intestinal microbial community structure in patients with esophageal cancer before and after chemotherapy.Short chain fatty acids-producing bacteria mainly on Lactobacillus and Rochella increases significantly in the intestinal microbiota of patients with clinical benefits.