Han Hye Seung, Lee Kyung-Yung, Lim So Dug, Kim Wan Seop, Hwang Tae Sook
Department of Pathology, Konkuk University School of Medicine, Seoul 143-729, Korea.
Oncol Lett. 2010 May;1(3):555-558. doi: 10.3892/ol_00000098. Epub 2010 May 1.
Helicobacter pylori (H. pylori) is a microaerophilic gram-negative bacterium known to be associated with chronic gastritis, peptic ulcer and gastric adenocarcinoma. In the present study, the presence of Helicobacter DNA was investigated using a Helicobacter species-specific 16S rRNA PCR amplification and pyrosequencing analysis in 51 resected gastric adenocarcinomas. DNA was extracted from paraffin-embedded tissues of resected gastric adenocarcinomas. PCR primers were designed to amplify the 133-bp PCR fragment in highly conserved regions of the 16S rRNA gene. The sequence of the PCR products was analyzed using a PSQ 96 system with SQA software. The pyrosequencing analysis of 16S rRNA showed that H. pylori was present in 47 (92.2%) of the 51 gastric adenocarcinomas. In the 4 H. pylori-negative cases, Helicobacter cinaedi (2 cases), Helicobacter mustelae (1 case) and Campylobacter hyointestinalis (1 case) were detected. Pyrosequencing technology was useful in the identification and differentiation of H. pylori from other species by analyzing the gene encoding 16S rRNA. Gastric adenocarcinoma tissues contain bacteria, and the majority are H. pylori. Helicobacter cinaedi, Helicobacter mustelae and Campylobacter hyointestinalis rarely occur. The roles of these organisms in the pathogenesis of gastric adenocarcinoma remain unclear.
幽门螺杆菌（H. pylori）是一种微需氧革兰氏阴性菌，已知与慢性胃炎、消化性溃疡和胃腺癌有关。在本研究中，采用幽门螺杆菌属特异性16S rRNA PCR扩增和焦磷酸测序分析，对51例切除的胃腺癌中幽门螺杆菌DNA的存在情况进行了研究。从切除的胃腺癌石蜡包埋组织中提取DNA。设计PCR引物以扩增16S rRNA基因高度保守区域中的133 bp PCR片段。使用带有SQA软件的PSQ 96系统分析PCR产物的序列。16S rRNA的焦磷酸测序分析表明，51例胃腺癌中有47例（92.2%）存在幽门螺杆菌。在4例幽门螺杆菌阴性病例中，检测到辛内西幽门螺杆菌（2例）、鼬幽门螺杆菌（1例）和猪肠弯曲菌（1例）。焦磷酸测序技术通过分析编码16S rRNA的基因，有助于从其他物种中鉴定和区分幽门螺杆菌。胃腺癌组织中含有细菌，大多数是幽门螺杆菌。辛内西幽门螺杆菌、鼬幽门螺杆菌和猪肠弯曲菌很少出现。这些微生物在胃腺癌发病机制中的作用仍不清楚。
