T-RFLP技术检测结直肠癌患者肠道黏膜相关菌群改变

目的研究结直肠癌患者肠道黏膜相关菌群组成差异,探索肠道菌群在结直肠癌发生发展中的作用。方法用末端限制片段长度多态性(Terminal restriction fragment length polymorphism,T-RFLP)技术分析50例结直肠癌患者癌组织、癌旁正常黏膜与健康对照组肠道黏膜相关细菌组成差异。结果与健康对照组相比,结直肠癌患者肠道黏膜相关细菌丰度显著增加(P0.05),多样性显著降低(P0.05)。结直肠癌患者癌组织与癌旁正常黏膜的黏膜相关细菌组成相近,但与健康对照组存在显著差异。MspI酶切的160bp、560bp的T-RF片段在结直肠癌患者癌组织及癌旁正常黏膜中为优势片段,而在健康对照组中缺失。相反,MspI酶切的66bp、74bp、141bp的T-RF片段在健康对照组为优势片段,但在结直肠癌癌组织及癌旁正常黏膜中缺失。结论肠道菌群失调与结直肠癌的发生发展密切相关。MspI酶切的66bp、74bp、141bp、160bp、560bp的T-RF片段所代表的细菌可能在结直肠癌的发生发展中起重要作用。

Change of intestinal mucosa-associated flora in colorectal cancer patients analyzed by T-RFLP

Abstract: ObjectiveTo analyze the composition of mucosa-associated flora in colorectal cancer patients and explore the role of intestinal microbiota in colorectal carcinogenesis. Methods 50 colorectal cancer patients and 46 healthy controls were enrolled in this study. Terminal restriction fragment length polymorphism (T-RFLP) was used to analyze the composition of mucosa-associated flora of cancerous tissue and noncancerous tissue in colorectal cancer patients vs health controls. Results Compared with healthy controls, the richness of mucosa-associated flora significantly increased while the diversity significantly decreased in colorectal cancer patients (P<0.05). The composition of mucosal microbiota of cancerous tissue was similar with that of noncancerous tissue in colorectal cancer patients, but both significantly differed from that of healthy controls. The 160 bp and 560 bp T-RFs digested by MspI were predominant in colorectal cancer patients but absent in healthy controls. On the contrast, the 66 bp, 74 bp and 141 bp T-RFs digested by MspI were predominant in healthy controls but absent in colorectal cancer patients. Conclusion Dysbiosis of mucosa-associated flora is closely related to colorectal cancer. The bacteria represented by 66 bp, 74 bp, 141 bp, 160 bp and 560 bp T-RFs (digested by MspI) may play an important role in colorectal carcinogenesis.