Fusobacterium nucleatum in gastroenterological cancer: Evaluation of measurement methods using quantitative polymerase chain reaction and a literature review

Yamamura,Kensuke; Baba,Yoshifumi; Miyake,Keisuke; Nakamura,Kenichi; Shigaki,Hironobu; Mima,Kosuke; Kurashige,Junji; Ishimoto,Takatsugu; Iwatsuki,Masaaki; Sakamoto,Yasuo; Yamashita,Yoichi; Yoshida,Naoya; Watanabe,Masayuki; Baba,Hideo

doi:10.3892/ol.2017.7001

Fusobacterium nucleatum in gastroenterological cancer: Evaluation of measurement methods using quantitative polymerase chain reaction and a literature review

Authors:
- Kensuke Yamamura
- Yoshifumi Baba
- Keisuke Miyake
- Kenichi Nakamura
- Hironobu Shigaki
- Kosuke Mima
- Junji Kurashige
- Takatsugu Ishimoto
- Masaaki Iwatsuki
- Yasuo Sakamoto
- Yoichi Yamashita
- Naoya Yoshida
- Masayuki Watanabe
- Hideo Baba
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Affiliations: Department of Gastroenterological Surgery, Graduate School of Medical Sciences, Kumamoto University, Kumamoto 860‑8556, Japan, Department of Gastroenterological Surgery, Cancer Institute Hospital, Japanese Foundation for Cancer Research, Tokyo 135‑8550, Japan
Published online on: September 19, 2017 https://doi.org/10.3892/ol.2017.7001
Pages: 6373-6378
Copyright: © Yamamura et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

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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