Rapid determination of AKAP12 promoter methylation levels in peripheral blood using methylation-sensitive high resolution melting (MS-HRM) analysis: Application in colorectal cancer期刊界 All Journals 搜尽天下杂志传播学术成果专业期刊搜索期刊信息化学术搜索

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Rapid determination of AKAP12 promoter methylation levels in peripheral blood using methylation-sensitive high resolution melting (MS-HRM) analysis: Application in colorectal cancer

Authors:	Weiwei Liu Ming Guan Bing Su Ji Li Weizhe Ma Chunfang Liu Min Li Yong Lin Yuan Lu

Institution:	1. Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, PR China;2. Central Laboratory, Huashan Hospital, Fudan University, Shanghai, PR China;3. Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York, USA;4. Department of Surgery, Huashan Hospital, Fudan University, Shanghai, PR China;1. Institute of Marine and Environmental Technology, University of Maryland Center for Environmental Science, Columbus Center, 701 E. Pratt Street, Baltimore, MD 21202, USA;2. College of Ocean and Earth Sciences, Xiamen University, Xiamen, Fujian 361102, China;1. Geisel School of Medicine, Hanover, NH, United States;2. Department of Psychiatry, Geisel School of Medicine, Hanover, NH, United States;3. Department of Psychiatry, Indiana University School of Medicine, Indianapolis, IN, United States;4. Departments of Radiology and Imaging Sciences, Indiana University School of Medicine, Indianapolis, IN, United States;5. Human Brain Collection Core, NIMH, Bethesda, MD, United States;3. Department of Orthopaedic Surgery and Graduate Program in Cell and Developmental Biology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107;4. Department of Orthopaedic Surgery, Keio University School of Medicine, Tokyo 108-8345, Japan;5. Department of Orthopaedic Surgery, Kitasato University Kitasato Institute Hospital, Tokyo 108-8641, Japan;1. Department of Biochemistry and Molecular Biology, Poznań University of Medical Sciences, 6 ?wi?cickiego St., 60-781 Poznań, Poland;2. First Department of Surgical Oncology and General Surgery, Wielkopolska Cancer Center, Poznań, Poland;3. Department of General Surgery, Oncologic Gastroenterological and Plastic Surgery, Poznań University of Medical Sciences, Poznań, Poland;1. Department of Gastroenterology, Translational Medicine Center, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, People''s Republic of China;2. BioChain (Beijing) Science and Technology, Inc., Beijing, People''s Republic of China;3. Department of Gastroenterology, Beijing Military General Hospital, Beijing, People''s Republic of China;4. Department of Endoscopy, Cancer Institute and Hospital, Chinese Academy of Medical Sciences, Beijing, People''s Republic of China;5. Department of Endoscopy, Beijing University Cancer Hospital and Institute, Beijing, People''s Republic of China

Abstract:	BackgroundColorectal cancer is the third most common form of cancer and hypermethylation has been shown to increase the risk of developing this disease. DNA hypermethylation in the A kinase anchor protein 12 (AKAP12/Gravin) promoter region and the accompanied underexpression of it has been noted in a variety of human cancers.MethodsWe applied methylation-specific high resolution melting (MS-HRM) technology to detect quantitatively A kinase anchor protein 12 (AKAP12/Gravin) methylation in peripheral blood from 100 colorectal cancer patients and 50 healthy volunteers and in 3 colorectal cancer cell lines.ResultsIn this study 48 of the 100 colorectal cancer samples (48%) were found to be methylated at the AKAP12 promoter region. AKAP12 methylation was significantly higher in the colorectal cancer samples with differentiation (p = 0.03). We also compared the results generated by MS-HRM with a traditional methylation-specific PCR (MSP) assay. We found that intra-assay variability ranged from 6.14 to 9.90% and inter-assay variability ranged from 14.5 to 17.2%. The AKAP12 MS-HRM assay was able to reproducibly detect 1% methylated DNA, whereas the MSP method was unable to detect less than 5% methylation.ConclusionsWe demonstrate the utility of quantitative AKAP12 MS-HRM analysis of promoter methylation in peripheral blood samples. AKAP12 MS-HRM quantitative methods with excellent detection capabilities have many promising applications in the research and diagnosis of colorectal cancer.

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