Analysis of fecal DNA methylation to detect gastrointestinal neoplasia.
BACKGROUND
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.


METHODS
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.


RESULTS
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).


CONCLUSIONS
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.
Nagasaka
Takeshi
T
Department of Gastroenterological Surgery and Surgical Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama City, Okayama 700-8558, Japan. takeshin@cc.okayama-u.ac.jp
Tanaka
Noriaki
N
Cullings
Harry M
HM
Sun
Dong-Sheng
DS
Sasamoto
Hiromi
H
Uchida
Takuyuki
T
Koi
Minoru
M
Nishida
Naoshi
N
Naomoto
Yoshio
Y
Boland
C Richard
CR
Matsubara
Nagahide
N
Goel
Ajay
A
D014408:Biomarkers, Tumor / Q000235:genetics; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D016001:Confidence Intervals; D019175:DNA Methylation; D018450:Disease Progression; D055088:Early Detection of Cancer; D016099:Endoscopy, Gastrointestinal; D005243:Feces; D005770:Gastrointestinal Neoplasms / Q000175:diagnosis / Q000235:genetics; D006801:Humans; D008403:Mass Screening / Q000379:methods; D008565:Membrane Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012372:ROC Curve; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D013447:Sulfites; D025521:Tumor Suppressor Proteins / Q000235:genetics*; D000094742:Secreted Frizzled-Related Proteins
D014408:Biomarkers, Tumor; D008565:Membrane Proteins; D013447:Sulfites; D025521:Tumor Suppressor Proteins; C425300:RASSF2 protein, human; C109618:SFRP2 protein, human; D000094742:Secreted Frizzled-Related Proteins; C042345:hydrogen sulfite
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