title: Methylation status of 5'-regulatory region of tumor necrosis factor alpha gene correlates with differentiation stages of monocytes.
abstract: The DNA methylation status of the 5'-regulatory region of human tumor necrosis factor-alpha (TNF-alpha) gene was examined using two human monocytic cell lines representing different differentiation stages: one cell line, THP-1, represents the most differentiated macrophage-like phenotypes and the other, HL-60, represents a least differentiated phenotype. Two restriction enzymes were used to detect methylated sites in the 5'-regulatory region of the TNF-alpha gene. The restriction enzyme Msp I, which recognizes both CCGG (unmethylated) and C(me)CGG (methylated) sequences, whereas Hpa II recognizes only the unmethylated CCGG sequence. Two Msp I sites located at -600 and +200 nucleotides, respectively, from the transcriptional initiation site were unmethylated in DNA from the THP-1, whereas these sites were methylated in the one from the HL-60. Treating the HL-60 cells with 1,25-vitamin D3, ( 1,25-(OH)2D3), which is known to induce monocytic cell differentiation, induce demethylation of the sites, suggesting the methylation status was correlated with the differentiation stages of monocytes.
authors.affiliation: Department of Medicine, Ceders-Sinai Medical Center, UCLA School of Medicine, California 90048, USA.
authors.forename: S
authors.initials: S
authors.lastname: Takei
authors.forename: D
authors.initials: D
authors.lastname: Fernandez
authors.forename: A
authors.initials: A
authors.lastname: Redford
authors.forename: H
authors.initials: H
authors.lastname: Toyoda
mesh_terms: D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D004247:DNA / Q000737:chemistry; D004273:DNA, Neoplasm / Q000737:chemistry; D019003:Deoxyribonuclease HpaII / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D008745:Methylation; D009000:Monocytes / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D013379:Substrate Specificity; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; D014409:Tumor Necrosis Factor-alpha; D004247:DNA; D019003:Deoxyribonuclease HpaII