结直肠腺瘤及结直肠癌患者肠道中梭杆菌属与产丁酸菌的定量研究

Quantitation of intestinal Fusobacterium and butyrateproducing bacteria in patients with colorectal adenomas and colorectal cancer

摘要:

摘要:【目的】通过观察梭杆菌属(Fusobacterium spp.)和两株产丁酸菌(Eubacterium rectale、Faecalibacterium prausnitzii)在结直肠癌患者及结直肠腺瘤患者粪便样品中的丰度差异，研究梭杆菌属和产丁酸菌数量变化在结直肠腺瘤和结直肠癌发生发展中的作用和意义。【方法】收集结直肠癌患者(n=19)、结直肠腺瘤患者(n=12)及健康人(n=19)3组粪便样品，提取细菌基因组DNA，利用实时荧光定量PCR技术定量检测3组样品中梭杆菌属(Fusobacterium spp.)、直肠真杆菌(Eubacterium rectale)、普拉梭菌(Faecalibacterium prausnitzii)以及总菌的16S rRNA基因的拷贝数，然后利用秩和检验两两比较3组样品中目标菌群的数量和丰度差异。【结果】结直肠癌组的梭杆菌属丰度显著高于结直肠腺瘤组(P=0.013)和健康组(P=0.000)，结直肠腺瘤组的梭杆菌属丰度显著高于健康组(P=0.002);结直肠腺瘤组普拉梭菌的丰度显著低于健康组(P=0.033) ;结直肠腺瘤组的总菌16S rRNA基因拷贝数也显著低于健康组(P=0.002);直肠真杆菌的水平在3组样品间没有显著差异。【结论】与健康人的粪便样品相比，结直肠腺瘤病人的粪便中产丁酸菌普拉梭菌数量下降，而结直肠腺瘤和结直肠癌病人的粪便样品中梭杆菌属数量增加;梭杆菌属和产丁酸菌数量上的变化提示它们可能与结直肠腺瘤和结直肠癌的发生密切相关。

关键词:关键词:结直肠癌，结直肠腺瘤，梭杆菌属，产丁酸菌，实时荧光定量PCR

Abstract:

Abstract:［Objective］To compare the abundance of 16S rRNA gene of intestinal Fusobacterium and butyrate-producing bacteria in patients with colorectal adenomas patients and colorectal cancer and to reveal the correlation between the target bacteria and the development of colorectal cancer．［Methods］Feces were collected from colorectal cancer patients (n=19)，colorectal adenomas patients (n= 12) and healthy subjects (n=19) ．Bacteria genome DNA from the fecal samples was used to quantitate the Fusobacterium，two butyrate-producing bacteria Eubacterium rectal，Faecalibacterium prausnitzii and total bacteria by real-time polymerase chain reaction． Then the variation of the target bacteria among different groups were assayed using Mann-Whitney U test．［Results］ The abundance of Fusobacterium was significantly higher in colorectal cancer patients than that in healthy subjects (P=0.000) and colorectal adenomas patients (P=0.013)，and it was significantly higher in colorectal cancer patients than that in colorectal adenomas patients (P=0.002)．F．prausnitzii was significantly lower in colorectal adenomas patients compared to healthy subjects (P=0.033)．The total bacteria count was significantly lower in the colorectal adenomas samples than that in the healthy samples (P=0.002)．There was no significantly difference of E．rectal between the three groups．［Conclusions］ The shifts in the colonic bacterial population may potentially contribute to the development of colorectal cancer．

Key words:Keywords: colorectal cancer，colorectal adenomas，Fusobacterium，butyrate-producing bacteria，real-time PCR