Fusobacterium nucleatum in gastroenterological cancer: Evaluation of measurement methods using quantitative polymerase chain reaction and a literature review.
Fusobacterium nucleatum in gastroenterological cancer: Evaluation of measurement methods using quantitative polymerase chain reaction and a literature review.
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DOI：
10.3892/ol.2017.7001
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发表时间：
2017-12
期刊：
影响因子：
2.9
通讯作者：
Baba H
中科院分区：
文献类型：
--
作者：
Yamamura K;Baba Y;Miyake K;Nakamura K;Shigaki H;Mima K;Kurashige J;Ishimoto T;Iwatsuki M;Sakamoto Y;Yamashita Y;Yoshida N;Watanabe M;Baba H
主要生活在口腔中的人类微生物群有核梭杆菌（Fusobacterium nucleatum）可引起牙周病，也与结直肠癌的发生有关。然而，核梭菌是否存在于其他胃肠癌组织中仍有待阐明。本研究评价了定量聚合酶链反应（qPCR）方法能否检测出食管、胃癌、胰腺癌和肝癌组织中具核梭菌的DNA，并测定其数量。使用从口腔细胞中提取的DNA，通过校准曲线确定qPCR检测的准确性。采用福尔马林固定石蜡包埋法（FFPE）对20例食管癌、胃癌、大肠癌、胰腺癌和肝癌患者的肿瘤组织和20例匹配的正常肿瘤组织进行核梭菌DNA含量测定。在qPCR检测中，核核梭菌和溶质载体有机阴离子转运蛋白家族成员2A1（参比样品）的周期阈值随输入DNA的数量呈线性下降（r2>0.99）。食管癌、胃癌和结直肠癌组织中具核梭菌的检出率分别为20%（4/20）、10%（2/20）和45%（9/20）。在肝癌和胰腺癌组织中未检出具核梭菌。冷冻组织和FFPE组织的qPCR结果一致。值得注意的是，与侵入区相比，浅表区检测到较高水平的具核梭菌。利用FFPE组织，采用qPCR方法对食管癌、胃癌和结直肠癌组织中具核梭菌进行检测。具核梭菌可能参与食管癌、胃癌和结直肠癌的发生。
The human microbiome Fusobacterium nucleatum, which primarily inhabits the oral cavity, causes periodontal disease and has also been implicated in the development of colorectal cancer. However, whether F. nucleatum is present in other gastroenterological cancer tissues remains to be elucidated. The present study evaluated whether quantitative polymerase chain reaction (qPCR) assays were able to detect F. nucleatum DNA and measure the quantity of F. nucleatum DNA in esophageal, gastric, pancreatic and liver cancer tissues. The accuracy of the qPCR assay was determined from a calibration curve using DNA extracted from cells from the oral cavity. Formalin-fixed paraffin-embedded (FFPE) tumor tissues from 20 patients with gastroenterological [esophageal (squamous cell carcinoma), gastric, colorectal, pancreatic and liver] cancer and 20 matched normal tissues were evaluated for F. nucleatum DNA content. The cycle threshold values in the qPCR assay for F. nucleatum and solute carrier organic anion transporter family member 2A1 (reference sample) decreased linearly with the quantity of input DNA (r2>0.99). The F. nucleatum detection rate in esophageal, gastric and colorectal cancer tissues were 20% (4/20), 10% (2/20) and 45% (9/20), respectively. F. nucleatum was not detected in liver and pancreatic cancer tissues. The qPCR results from the frozen and FFPE tissues were consistent. Notably, F. nucleatum was detected at a higher level in superficial areas compared with the invasive areas. F. nucleatum in esophageal, gastric and colorectal cancer tissues was evaluated by qPCR using FFPE tissues. F. nucleatum may be involved in the development of esophageal, gastric and colorectal cancer.
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