BARHL2 Methylation Using Gastric Wash DNA or Gastric Juice Exosomal DNA is a Useful Marker For Early Detection of Gastric Cancer in an H. pylori-Independent Manner.
BARHL2 Methylation Using Gastric Wash DNA or Gastric Juice Exosomal DNA is a Useful Marker For Early Detection of Gastric Cancer in an H. pylori-Independent Manner.
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DOI：
10.1038/ctg.2016.40
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发表时间：
2016-07-21
影响因子：
3.6
通讯作者：
Itoh F
中科院分区：
文献类型：
--
作者：
Yamamoto H;Watanabe Y;Oikawa R;Morita R;Yoshida Y;Maehata T;Yasuda H;Itoh F
本研究的主要目的是利用胃洗液衍生的 DNA 和/或胃液衍生的外泌体 DNA (exoDNA) 开发甲基化分析流程，并评估其在临床环境中早期检测胃癌 (GC) 的适用性。我们分析了 GC 细胞系和组织中 BarH 样 2 同源盒蛋白 (BARHL2) 的变化，以及从 128 次胃洗液和 30 次胃液衍生的外泌体中获得的 DNA 的变化。用编码 BARHL2 的质粒转染 GC 细胞系，并进行增殖、集落形成和基因表达分析。在七个 GC 细胞系中的三个中检测到高水平的 BARHL2 甲基化；与此一致的是，这些细胞系表达低水平的 BARHL2。用 5-aza-2'-脱氧胞苷处理这些细胞系可恢复 BARHL2 表达。无论是否感染幽门螺杆菌，18 份正常样本和 14 份萎缩性胃炎样本中的 BARHL2 甲基化水平均较低。内镜切除 (ER) 前早期 GC 患者的胃洗液衍生 DNA 中观察到高水平的 BARHL2 甲基化，但治疗性 ER 后甲基化显着降低。使用胃液来源的外切 DNA 样本进行分析表明，BARHL2 甲基化的曲线下面积为 0.923，在区分 GC 患者和非 GC 对照方面具有 90% 的敏感性和 100% 的特异性。在所有正常胃上皮细胞以及胃炎和腺瘤患者的细胞中都发现了 BARHL2 核免疫反应性。相反，在绝大多数GC组织中观察到BARHL2表达缺失。最后，将 BARHL2 转染至 MKN7 和 MKN45 细胞系可显着抑制其增殖和形成集落的能力。使用胃洗液衍生的 DNA 和/或胃液衍生的外切 DNA 对 BARHL2 进行甲基化分析，可用于临床环境中早期检测 GC。
The main purpose of this study was to develop a methylation analysis pipeline by using gastric wash-derived DNA and/or gastric juice-derived exosomal DNA (exoDNA), and to evaluate its suitability for the early detection of gastric cancer (GC) in clinical settings. We analyzed alterations of BarH-like 2 homeobox protein (BARHL2) in GC cell lines and tissues, as well as in DNA obtained from 128 gastric washes and 30 gastric juice-derived exosomes. GC cell lines were transfected with plasmids encoding BARHL2 and subjected to proliferation, colony formation, and gene expression analyses. High levels of BARHL2 methylation were detected in three of seven GC cell lines; consistent with this, these cell lines expressed low levels of BARHL2. Treatment of these cell lines with 5-aza-2′-deoxycytidine restored BARHL2 expression. Levels of BARHL2 methylation in 18 normal and 14 atrophic gastritis samples were low irrespective of Helicobacter pylori infection. High levels of BARHL2 methylation were observed in gastric wash-derived DNA obtained from early GC patients before endoscopic resection (ER), but methylation was significantly lower after curative ER. Analysis using gastric juice-derived exoDNA samples revealed that BARHL2 methylation yielded an area under the curve of 0.923 with 90% sensitivity and 100% specificity with respect to discriminating GC patients from non-GC controls. BARHL2 nuclear immunoreactivity was found in all normal gastric epithelial cells and in cells from patients with gastritis and adenoma. In contrast, loss of BARHL2 expression was observed in the vast majority of the GC tissues. Finally, transfection of BARHL2 into MKN7 and MKN45 cell lines significantly inhibited their proliferation and ability to form colonies. Methylation analysis of BARHL2 using gastric wash-derived DNA and/or gastric juice-derived exoDNA could be useful for early detection of GC in clinical settings.
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影响因子：
29.4
作者：
Watanabe Y;Kim HS;Castoro RJ;Chung W;Estecio MR;Kondo K;Guo Y;Ahmed SS;Toyota M;Itoh F;Suk KT;Cho MY;Shen L;Jelinek J;Issa JP
通讯作者：
Issa JP
DOI：
10.1073/pnas.1014017108
发表时间：
2011-02-08
影响因子：
11.1
作者：
Juraver-Geslin, Hugo A.;Ausseil, Jerome J.;Durand, Beatrice C.
通讯作者：
Durand, Beatrice C.
影响因子：
6.3
作者：
Nakajima, Takeshi;Enomoto, Shotaro;Ushijima, Toshikazu
通讯作者：
Ushijima, Toshikazu
影响因子：
6.4
作者：
Shin, Cheol Min;Kim, Nayoung;Jung, Hyun Chae
通讯作者：
Jung, Hyun Chae
影响因子：
3.2
作者：
Baba, Satoshi;Oishi, Yoshichika;Itoh, Fumio
通讯作者：
Itoh, Fumio