Depletion of Embryonic Stem Cell Signature by Histone Deacetylase Inhibitor in NCCIT Cells: Involvement of Nanog Suppression

You, Jueng Soo; Kang, Jaeku; Seo, Dong Wan; Park, Jae Hyun; Park, Jong Woo; Lee, Jae Cheol; Jeon, Yae Jee; Cho, Eun Jung; Han, Jeung-Whan

doi:10.1158/0008-5472.can-08-4953

Cited by 50 publications

(56 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To investigate the roles of distinct epigenetic mechanisms in silencing pluripotency genes, we used embryonic carcinoma cell line NCCIT (19,20), colon cancer cell line HCT116, and cell line DKO1, the hypomethylated derivative of HCT116 that exhibits 95% reduced DNA methylation because of the genetic knockdown of DNA methyltransferases DNMT3B and DNMT1 (21,22). These cell lines were selected because they represent distinct transcriptional contexts of pluripotency genes in the presence and absence of DNA methylation.…”

Section: Resultsmentioning

confidence: 99%

OCT4 establishes and maintains nucleosome-depleted regions that provide additional layers of epigenetic regulation of its target genes

You

Kelly²,

Carvalho³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

118

View full text Add to dashboard Cite

Recent epigenome-wide mapping studies describe nucleosomedepleted regions (NDRs) at transcription start sites and enhancers. However, these static maps do not address causality or the roles of NDRs in gene control, and their relationship to transcription factors and DNA methylation is not well understood. Using a highresolution single-molecule mapping approach to simultaneously investigate endogenous DNA methylation and nucleosome occupancies on individual DNA molecules, we show that the unmethylated OCT4 distal enhancer has an NDR, whereas NANOG has a clear NDR at its proximal promoter. These NDRs are maintained by binding of OCT4 and are required for OCT4 and NANOG expression. Differentiation causes a rapid loss of both NDRs accompanied by nucleosome occupancy, which precedes de novo DNA methylation. NDRs can be restored by forced expression of OCT4 in somatic cells but only when there is no cytosine methylation. These data show the central role of the NDRs, established by OCT4, in ensuring the autoregulatory loop of pluripotency and, furthermore, that de novo methylation follows the loss of NDRs and stabilizes the suppressed state. D uring development, each cell acquires its own epigenetic signature that provides guidelines to its cellular identity (1, 2). This epigenetic signature is accomplished by multiple epigenetic mechanisms, including DNA methylation, histone modifications, nucleosome positioning, and noncoding RNAs (3, 4). Pluripotent cells have a distinctive signature that is more dynamic compared with differentiated cells and allows for selfrenewal and pluripotency. Developmentally important genes are bivalent in embryonic stem cells, containing both active and repressive histone modifications (5, 6). The transcription factors OCT4, SOX2, and NANOG are known as core regulators of the transcription circuitry in pluripotent cells. The transcription autoregulatory loop ensures high levels of expression of these key stemness genes; they bind to their own regulatory regions, thereby maintaining expression patterns necessary for establishing and preserving pluripotent states (7).The nucleosome is the basic unit of chromatin and consists of DNA wound around a histone octamer protein core to achieve high compaction. Besides its role in packing the genome, pioneering studies have shown that nucleosome occupancy at gene promoters inhibits transcription initiation (8) and plays a critical role in epigenetic regulation (9). Genome-wide studies have shown that nucleosome-depleted regions (NDRs) are present at the transcription start sites of active genes and enhancers (10-14). More recently, genome-wide studies have begun to focus on the relationship between nucleosome positioning and gene expression (15, 16). Despite these observations, the role of dynamic nucleosome occupancy at gene regulatory regions and the relationship with transcription factors has not been well characterized at high resolution, particularly during the initial steps when embryonic stem cells lose their pluripotency.In this study, we used ...

show abstract

Section: Resultsmentioning

confidence: 99%

OCT4 establishes and maintains nucleosome-depleted regions that provide additional layers of epigenetic regulation of its target genes

You

Kelly²,

Carvalho³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

118

View full text Add to dashboard Cite

show abstract

“…NANOG expression in embryo development is crucial and probably depends on its ability to regulate the expression of OCT4 and SOX2, which have been implicated in transcription regulation of a number of genes in the pluripotency network [20,63]. It was also demonstrated that HDAC1 mediated suppression of OCT4 and SOX2 results in suppression of NANOG [64]. It has been suggested that HDAC1, OCT4 and NANOG form the NODE complex responsible for regulation of gene expression and for maintaining pluripotency [65].…”

Section: Discussionmentioning

confidence: 99%

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

Gibence

Brahmasani

Mahesh

et al. 2014

J Assist Reprod Genet

View full text Add to dashboard Cite

Purpose The levels and timing of expression of genes like BCLXL, HDAC1 and pluripotency marker genes namely, OCT4, SOX2, NANOG and KLF4 are known to influence preimplantation embryo development. Despite this information, precise understanding of their influence during preimplantation embryo development is lacking. The present study attempts to compare the expression of these genes in the in vivo and in vitro developed preimplantation embryos. Methods The in vivo and in vitro developed rabbit embryos collected at distinct developmental stages namely, pronuclear, 2 cell, 4 cell, 8 cell, 16 cell, Morula and blastocyst were compared at the transcriptional and translational levels using Real Time PCR and immunocytochemical studies respectively. Results The study establishes the altered levels of candidate genes at the transcriptional level and translational level with reference to the zygotic genome activation (ZGA) phase of embryo development in the in vivo and in vitro developed embryos. The expression of OCT4, KLF4, NANOG and SOX2 genes were higher in the in vitro developed embryos whereas and HDAC1 was lower. BCLXL expression had its peak at ZGA in in vivo developed embryos. Protein expression of all the candidate genes was observed in the embryos. BCLXL, KLF4 and NANOG exhibited diffused localisation whereas HDAC1, OCT4, and SOX2 exhibited nuclear localisation. Conclusions This study leads to conclude that BCLXL peak expression at the ZGA phase may be a requirement for embryo development. Further expression of all the candidate genes was influenced by ZGA phase of development at the transcript level, but not at the protein level.

show abstract

“…This assay is considered the most stringent in vitro test for determining malignant transformation. 53 It has been shown that the ability of tumor cells to form (10 7 pfu) IV on the indicated day. Lungs were removed on day 12 following tumor challenge, at which time metastases were macroscopically visible (a).…”

Section: Resultsmentioning

confidence: 99%

Dasatinib alters the metastatic phenotype of B16-OVA melanoma in vivo

Fraser

Lousberg

Guerin

et al. 2010

Cancer Biology & Therapy

View full text Add to dashboard Cite

Depletion of Embryonic Stem Cell Signature by Histone Deacetylase Inhibitor in NCCIT Cells: Involvement of Nanog Suppression

Cited by 50 publications

References 37 publications

OCT4 establishes and maintains nucleosome-depleted regions that provide additional layers of epigenetic regulation of its target genes

OCT4 establishes and maintains nucleosome-depleted regions that provide additional layers of epigenetic regulation of its target genes

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

Dasatinib alters the metastatic phenotype of B16-OVA melanoma in vivo

Contact Info

Product

Resources

About