Demethylating agent, 5-azacytidine, reverses differentiation of embryonic stem cells

Tsuji-Takayama, Kazue; Inoue, Toshiya; Ijiri, Yoshihiro; Otani, Takeshi; Motoda, Ryuichi; Nakamura, Shuji; Orita, Kunzo

doi:10.1016/j.bbrc.2004.08.052

Cited by 68 publications

(48 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Oct4 expression is down-regulated as mESCs differentiate by increased promoter methylation. An alteration of DNA methylation and histone modification by 5-aza-2-deoxycytidine or trichostatin A (TSA) at the Oct4 promoter lead to failed stem cell differentiation, which suggests that epigenetics plays an intricate role in stem cell regulation (Tsuji-Takayama et al, 2004).…”

Section: Cdk2ap1 In Epigenetic Regulationmentioning

confidence: 99%

Double Edge

et al. 2011

View full text Add to dashboard Cite

Cancer research has been devoted toward an understanding of the molecular regulation and functional significance of cell-cycle regulators in the pathogenesis and development of cancers. Cyclin-dependent Kinase 2-associated Protein 1 (CDK2AP1) is one such cell-cycle regulator, originally identified as a growth suppressor and a prognostic marker for human oral/head and neck cancers. Functional importance and the molecular mechanism of CDK2AP1-mediated cell-cycle regulation have been documented over the years. Recent progress has shown that CDK2AP1 is a competency factor in embryonic stem cell differentiation. Deletion of CDK2AP1 leads to early embryonic lethality, potentially through altered differentiation capability of embryonic stem cells. More intriguingly, CDK2AP1 exerts its effect on stem cell maintenance/differentiation through epigenetic regulation. Cancer cells and stem cells share common cellular characteristics, most prominently in maintaining high proliferative potential through an unconventional cell-cycle regulatory mechanism. Cross-talk between cellular processes and molecular signaling pathways is frequent in any biological system. Currently, it remains largely elusive how cell-cycle regulation is mechanistically linked to epigenetic control. Understanding the molecular mechanism underlying CDK2AP1-mediated cell-cycle regulation and epigenetic control will set an example for establishing a novel and effective molecular link between these two important regulatory mechanisms.

show abstract

Section: Cdk2ap1 In Epigenetic Regulationmentioning

confidence: 99%

Double Edge

et al. 2011

View full text Add to dashboard Cite

show abstract

“…All firefly luciferase activity was normalized to Renilla luciferase. described previously (30). Briefly, the genomic DNA (5 g) was digested with EcoRI and then incubated in 3.2 M sodium bisulfite (Sigma-Aldrich) and 500 nM hydroquinone (Sigma-Aldrich) for 16 h at 55°C and then desalted using a DNA cleanup column (Promega).…”

Section: Transient Transfection and Reporter Gene Assaysmentioning

confidence: 99%

The Production of IL-10 by Human Regulatory T Cells Is Enhanced by IL-2 through a STAT5-Responsive Intronic Enhancer in the IL-10 Locus

Tsuji-Takayama

Suzuki

Yamamoto

et al. 2008

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

STAT5 molecules are key components of the IL-2 signaling pathway, the deficiency of which often results in autoimmune pathology due to a reduced number of CD4+CD25+ naturally occurring regulatory T (Treg) cells. One of the consequences of the IL-2-STAT5 signaling axis is up-regulation of FOXP3, a master control gene for naturally occurring Treg cells. However, the roles of STAT5 in other Treg subsets have not yet been elucidated. We recently demonstrated that IL-2 enhanced IL-10 production through STAT5 activation. This occurred in two types of human Treg cells: a novel type of umbilical cord blood-derived Treg cell, termed HOZOT, and Tr1-like Treg cells, IL-10-Treg. In this study, we examined the regulatory mechanisms of IL-10 production in these Treg cells, focusing specifically on the roles of STAT5. By performing bioinformatic analysis on the IL-10 locus, we identified one STAT-responsive element within intron 4, designated I-SRE-4, as an interspecies-conserved sequence. We found that I-SRE-4 acted as an enhancer element, and clustered CpGs around the I-SRE-4 were hypomethylated in IL-10-producing Treg cells, but not in other T cells. A gel-shift analysis using a nuclear extract from IL-2-stimulated HOZOT confirmed that CpG DNA methylation around I-SRE-4 reduced STAT5 binding to the element. Chromatin immunoprecipitation analysis revealed the in situ binding of IL-2-activated STAT5 to I-SRE-4. Thus, we provide molecular evidence for the involvement of an IL-2-STAT5 signaling axis in the expression of IL-10 by human Treg cells, an axis that is regulated by the intronic enhancer, I-SRE-4, and epigenetic modification of this element.

show abstract

“…Moreover, Hattori et al (11) demonstrated methylation of the mouse Oct-4 promoter region in trophoblast stem cells. In an attempt to study the possibility of reversing the differentiation process, Tsuji-Takayama et al (12) treated differentiated ES cells with a demethylating agent and found an increase in the expression of ES-specific genes such as Oct-4, Nanog, and Sox2. However, in this case, expression of any of these three genes could be an indirect consequence of demethylation of another gene.…”

mentioning

confidence: 99%

Sequential DNA Methylation of the Nanog and Oct-4 Upstream Regions in Human NT2 Cells during Neuronal Differentiation

Deb-Rinker

Jezierski

et al. 2005

Journal of Biological Chemistry

169

124

View full text Add to dashboard Cite

Human NT2 cells, which differentiate into neurons and astrocytes, initially express and then permanently down-regulate Nanog and Oct-4 (POU5F1). We investigated the relationship between the expression of these genes and the methylation state of their 5-flanking regions. Gene expression and DNA methylation were assayed with quantitative polymerase chain reaction and bisulfite genomic sequencing, respectively. Retinoic acid-induced differentiation of NT2 cells to neurons is accompanied by a sequential decrease in the expression of both genes, paralleled by sequential epigenetic modification of their upstream regions. This is the first report demonstrating changes in DNA methylation in the promoter regions of Nanog and Oct-4 in a human cell line.

show abstract

Demethylating agent, 5-azacytidine, reverses differentiation of embryonic stem cells

Cited by 68 publications

References 37 publications

Double Edge

Double Edge

The Production of IL-10 by Human Regulatory T Cells Is Enhanced by IL-2 through a STAT5-Responsive Intronic Enhancer in the IL-10 Locus

Sequential DNA Methylation of the Nanog and Oct-4 Upstream Regions in Human NT2 Cells during Neuronal Differentiation

Contact Info

Product

Resources

About