Analysis of Fecal DNA Methylation to Detect Gastrointestinal Neoplasia
Analysis of Fecal DNA Methylation to Detect Gastrointestinal Neoplasia
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DOI：
10.1093/jnci/djp265
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发表时间：
2009-09-16
影响因子：
10.3
通讯作者：
Goel, Ajay
中科院分区：
文献类型：
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作者：
Nagasaka, Takeshi;Tanaka, Noriaki;Goel, Ajay
无创筛查试验的发展对降低胃肠道肿瘤的死亡率非常重要。我们试图通过分析粪便中脱落的癌细胞的DNA甲基化来开发这样一种测试。我们首先分析了来自788个原发胃和结直肠组织标本的RASSF2和SFRP2基因启动子的甲基化，以确定甲基化模式是否可以作为胃肠道肿瘤发生的分期依赖性生物标志物。接下来，我们开发了一种新的策略，使用亚硫酸氢钠和荧光聚合酶链反应方法对DNA进行单步修饰，以测量粪便DNA中的异常甲基化。我们分析了296份来自不同患者的粪便样本中RASSF2和SFRP2启动子的甲基化，其中包括21例胃肿瘤患者，152例结直肠肿瘤患者，10例胃肠道管腔非肿瘤性或炎性病变患者。来自组织的DNA分析显示，仅在晚期胃和结直肠肿瘤中，这两个基因启动子存在广泛的甲基化。该方法在57.1%的胃癌患者、75.0%的结直肠癌患者和44.4%的晚期结直肠腺瘤患者的粪便DNA中成功鉴定出一种或多种甲基化标记物，但在无肿瘤或活动性疾病的受试者中仅为10.6%(差异，胃癌vs未患病= 46.5%，95%置信区间(CI) = 24.6% ~ 68.4%, P < 0.001；差异，结直肠癌vs未患病= 64.4%,95% CI = 53.5% ~ 75.2%, P < 0.001；结直肠腺瘤vs未病变= 33.8%,95% CI = 14.2% ~ 53.4%, P < 0.001)。相对于非肿瘤性疾病，粪便DNA中RASSF2和SFRP2启动子的甲基化与胃肠道肿瘤的存在相关。我们的新型粪便DNA甲基化试验提供了一种可能的手段，不仅对结直肠肿瘤，而且对胃肿瘤的无创筛查。
The development of noninvasive screening tests is important to reduce mortality from gastrointestinal neoplasia. We sought to develop such a test by analysis of DNA methylation from exfoliated cancer cells in feces.We first analyzed methylation of the RASSF2 and SFRP2 gene promoters from 788 primary gastric and colorectal tissue specimens to determine whether methylation patterns could act as stage-dependent biomarkers of gastrointestinal tumorigenesis. Next, we developed a novel strategy that uses single-step modification of DNA with sodium bisulfite and fluorescence polymerase chain reaction methodology to measure aberrant methylation in fecal DNA. Methylation of the RASSF2 and SFRP2 promoters was analyzed in 296 fecal samples obtained from a variety of patients, including 21 with gastric tumors, 152 with colorectal tumors, and 10 with non-neoplastic or inflammatory lesions in the gastrointestinal lumen.Analysis of DNA from tissues showed presence of extensive methylation in both gene promoters exclusively in advanced gastric and colorectal tumors. The assay successfully identified one or more methylated markers in fecal DNA from 57.1% of patients with gastric cancer, 75.0% of patients with colorectal cancer, and 44.4% of patients with advanced colorectal adenomas, but only 10.6% of subjects without neoplastic or active diseases (difference, gastric cancer vs undiseased = 46.5%, 95% confidence interval (CI) = 24.6% to 68.4%, P < .001; difference, colorectal cancer vs undiseased = 64.4%, 95% CI = 53.5% to 75.2%, P < .001; difference, colorectal adenoma vs undiseased = 33.8%, 95% CI = 14.2% to 53.4%, P < .001).Methylation of the RASSF2 and SFRP2 promoters in fecal DNA is associated with the presence of gastrointestinal tumors relative to non-neoplastic conditions. Our novel fecal DNA methylation assay provides a possible means to noninvasively screen not only for colorectal tumors but also for gastric tumors.