期刊
CLINICAL COLORECTAL CANCER
卷 9, 期 3, 页码 168-176出版社
CIG MEDIA GROUP, LP
DOI: 10.3816/CCC.2010.n.023
关键词
CpG islands; Melting curve analyses; Noninvasive diagnosis; PCR
类别
资金
- Spanish Ministry of Education and Science [AGL2004-07579-04, SAF 06-6084]
- Fondo de Investigacion Sanitaria [FIS 07/0475]
- Indas Biotech, SLU [PI 0005]
- Fundacio Gastroenterologia Dr. Francisco Vilardell [F05-01]
- Accion en Cancer (Instituto de Salud Carlos III) [RD06/0020/1050, RD06/0020/1051]
- Accion Transversal contra el Cancer
Background: Previous studies showed that the assessment of promoter hypermethylation of a limited number of genes in tumor biopsies may identify the majority of colorectal tumors. This study aimed to assess the clinical usefulness of a panel of methylation biomarkers in stool DNA in the identification of colorectal tumors, using methylation-specific melting curve analysis (MS-MCA), a technique that simultaneously analyzes all cytosine-phosphate-guanine (CpG) residues within a promoter. Materials and Methods: The promoter methylation status of 4 tumor-related genes (RARB2, p16(INK4a), MGMT, and APC) was analyzed in DNA stool samples and corresponding tissues in an initial set of 12 patients with newly diagnosed primary colorectal carcinomas and 20 patients with newly diagnosed colorectal adenomas, using methylation-specific polymerase chain reaction. Results were replicated in a set of 82 patients (20 healthy subjects, 16 patients with inflammatory bowel disease (IBD), 20 patients with adenomas, and 26 patients with carcinomas), using MS-MCA analyses. Results: In the initial set, >= 1 positive methylation marker was detected in the stools of 9 of 12 patients (75%) with carcinomas and 12 of 20 patients (60%) with adenomas, with no false-positive results. Stool analyses missed 7 methylated lesions (25%). In the replication set, stool DNA testing detected 16 of 26 carcinomas (62%) and 8 of 20 adenomas (40%). The MS-MCAs missed 14 methylated tumors (37%). No aberrant methylation was evident in healthy subjects, but the RARB2 marker was positive in 2 of 15 stool samples (13%) of patients with IBD. Conclusion: Analysis via MS-MCA of a panel of methylation markers in stool DNA may offer a good alternative in the early, noninvasive detection of colorectal tumors.
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