Semi-quantitative detection of GADD45-gamma methylation levels in gastric, colorectal and pancreatic cancers using methylation-sensitive high-resolution melting analysis

• Published in: Journal of cancer research and clinical oncology Vol. 136; no. 8; pp. 1267 - 1273
• Main Authors: Zhang, Wei, Li, Tianrun, Shao, Yong, Zhang, Chao, Wu, Qi, Yang, Hong, Zhang, Jie, Guan, Ming, Yu, Bo, Wan, Jun
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.08.2010; Springer-Verlag; Springer; Springer Nature B.V
• Subjects: Antineoplastic agents; Biological and medical sciences; Cancer; Cancer Research; colorectal neoplasms; Colorectal Neoplasms - genetics; Colorectal Neoplasms - pathology; DNA Damage; DNA Methylation; DNA Primers; DNA, Neoplasm - genetics; Epigenesis, Genetic; Epigenetics; GADD45G; Gastroenterology. Liver. Pancreas. Abdomen; Genetic Variation; Hematology; HRM; Humans; Internal Medicine; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Liver. Biliary tract. Portal circulation. Exocrine pancreas; Medical sciences; Medicine; Medicine & Public Health; Methylation; Neoplasm Staging; Oncology; Original Paper; Pancreatic cancer; Pancreatic Neoplasms - genetics; Pancreatic Neoplasms - pathology; Pharmacology. Drug treatments; Promoter Regions, Genetic; Proteins; Reference Values; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm - genetics; stomach neoplasms; Stomach Neoplasms - genetics; Stomach Neoplasms - pathology; Stomach. Duodenum. Small intestine. Colon. Rectum. Anus; Tumors; HRM; Methylation; Gastric cancer; Pancreatic cancer; Colorectal cancer; GADD45G; Stomach; High resolution; Rectal disease; Digestive system; Malignant tumor; Stomach cancer; Colonic disease; Pancreas cancer; Digestive diseases; Intestinal disease; Diagnosis; Detection; Cancer; Gastric disease; Pancreatic disease; Quantitative analysis
• ISSN: 0171-5216; 1432-1335
• DOI: 10.1007/s00432-010-0777-z

Purpose GADD45 is a family of proteins involved in DNA damage response and cell growth arrest. GADD45G was identified as an interleukin-2-induced immediate-early gene, and methylation of GADD45G was studied in various tumor cell lines and a few primary tumor samples. High-resolution melting (HRM) analysis has been used as a novel tool for analysis of promoter methylation. Methods In our study, we used HRM analysis to detect the methylation levels of GADD45G gene in 100 gastric cancers, 100 colorectal cancers, 70 pancreatic cancers and equal number of adjacent normal tissues. Results The frequency of GADD45G methylation in all three types of cancers was significantly higher than that in normal tissues. Consistent with previous reports, expression levels of GADD45G were inversely correlated with methylation levels. But we did not find significant association between GADD45G methylation status and TNM staging in all three types of cancers. Conclusions In summary, application of HRM analysis to large amount of clinical samples proves to be a fast and high-throughput way to investigate the epigenetic status of GADD45G. And this is the first study to evaluate the prevalence of GADD45G methylation based on large amount of tumor samples, showing that epigenetic regulation of GADD45G was associated with carcinogenesis.