title: Rapid assessment of the heterogeneous methylation status of CEBPA in patients with acute myeloid leukemia by using high-resolution melting profile.
abstract: Epigenetic inactivation of tumor-suppressor genes, often in association with aberrant DNA methylation of CpG islands in the promoter region of these genes, is a key factor in tumorigenesis. CCAAT/enhancer binding protein alpha (CEBPA) methylation is a favorable prognostic biomarker for acute myeloid leukemia; however, rather than the complete methylation observed in inherited disorders, CEBPA methylation is heterogeneous. In this study, we established an algorithm called the "methylation index," deduced from high-resolution melting profiles, which includes Tm shifting (ΔTm) and Tm width ratio (fold of width), to evaluate the heterogeneous methylation status. The methylation index was highly correlated with the exact methylation levels detected by using the MassARRAY method (R(2) = 0.80; P < 0.001). Within-run reproducibility for the methylation index was 0.9% as the coefficient of variation, and between-run reproducibility was 2.6%. It was determined that with a cutoff methylation index of 1.412, the best measures of sensitivity and specificity could be obtained (97.14% and 95.89%, respectively) to discern low or high CEBPA methylation status. This novel algorithm for calculation of the methylation index from high-resolution melting profiles for CEBPA methylation is compatible with measurement of the methylation level as assayed using MassARRAY and could be a simple and efficient screening method for determination of CEBPA methylation status in acute myeloid leukemia.
authors: Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University, Taipei, Taiwan.
authors: Tsung-Chin
authors: TC
authors: Lin
authors: Sin-Sien
authors: SS
authors: Jiang
authors: Wen-Chien
authors: WC
authors: Chou
authors: Hsin-An
authors: HA
authors: Hou
authors: Yu-Min
authors: YM
authors: Lin
authors: Chia-Ling
authors: CL
authors: Chang
authors: Cherng-An
authors: CA
authors: Hsu
authors: Hwei-Fang
authors: HF
authors: Tien
authors: Liang-In
authors: LI
authors: Lin
mesh_terms: D000465:Algorithms; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics*; D045744:Cell Line, Tumor; D019175:DNA Methylation / Q000235:genetics*; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000473:pathology; D009367:Neoplasm Staging; D009691:Nucleic Acid Denaturation / Q000235:genetics*; D044366:Transition Temperature
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha