Dynamic DNA methylation change in the CpG island region of p15 during human myeloid development.
We examined the kenetics of p15 methylation and expression during myeloid development. We treated human cord blood CD34+ cells with either GM-CSF alone or in combination with stem cell factor and followed methylation at this locus using bisulfite genomic sequencing. CD34+ cells were found to be either fully methylated or completely unmethylated at 27 CpG dinucleotide sites in exon 1 and at 18 CpG sites in the promoter region of the p15 gene. A time-course study showed that the percentage of the allelic methylation of p15 CpG island increased to approximately 50% to 60% until 7 days after cytokine stimulation, then decreased to less than 10% after 21 days. The methylation was also observed in bone marrow CD34+ cells exposed to GM-CSF. p15 expression varied inversely with methylation. Expression was negligible or at low levels until 14 days, after which it increased substantially. The frequency of myeloid colony-forming cells in the progeny decreased and myeloid-specific markers increased in the later stages. Based on our observations on cells grown with GM-CSF and 5-aza-2'-deoxycytidine, DNA methylation of the p15 promoter region CpG island appears to be associated with proliferation rather than differentiation of normal human myeloid progenitors.
Sakashita
K
K
Department of Pediatrics, Institute of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University School of Medicine, 3-1-1, Asahi, Matsumoto, 390-8621, Japan.
Koike
K
K
Kinoshita
T
T
Shiohara
M
M
Kamijo
T
T
Taniguchi
S
S
Kubota
T
T
D000483:Alleles; D018952:Antigens, CD34 / Q000378:metabolism; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics*; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D018899:CpG Islands; D050762:Cyclin-Dependent Kinase Inhibitor p15; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D004247:DNA / Q000235:genetics; D019175:DNA Methylation; D005312:Fetal Blood / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D016147:Genes, Tumor Suppressor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006098:Granulocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D006410:Hematopoiesis / Q000235:genetics* / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D019089:Stem Cell Factor / Q000494:pharmacology; D025521:Tumor Suppressor Proteins
D018952:Antigens, CD34; C493966:CDKN2B protein, human; D018797:Cell Cycle Proteins; D050762:Cyclin-Dependent Kinase Inhibitor p15; D019941:Cyclin-Dependent Kinase Inhibitor p16; D019089:Stem Cell Factor; D025521:Tumor Suppressor Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA