Short hairpin RNA-directed cytosine (CpG) methylation of the RASSF1A gene promoter in HeLa cells

Castanotto, Daniela; Tommasi, Stefania; Li, Mingjie; Li, Haitang; Yanow, Stephanie K.; Pfeifer, Gerd P.; Rossi, John J.

doi:10.1016/j.ymthe.2005.03.003

Cited by 137 publications

(115 citation statements)

References 18 publications

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“…H3K9me3 was detected adjacent to the target region and we also found significant increase in its levels promoter-targeting siRNA-treated cells, which suggests that histone modification participates in gene silencing of HR-HPV E6 and E7 at the transcriptional level. Several publications also demonstrated the consistency between transcriptional gene silencing and DNA methylation in human cells (Morris et al, 2004;Castanotto et al, 2005). In our result, we found no methylation in the promoter of E6 and E7 with pyrosequencing.…”

Section: E6 E7supporting

confidence: 78%

Gene silencing of HPV16 E6/E7 induced by promoter-targeting siRNA in SiHa cells

Hong

Lü

et al. 2009

Br J Cancer

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BACKGROUND: Recently, transcriptional gene silencing induced by small interfering RNA (siRNA) was found in mammalian and human cells. However, previous studies focused on endogenous genes. METHODS: In this study, we designed siRNA targeting the promoter of human papillomavirus 16 E6/E7 and transfected it into the cervical cancer cell line, SiHa. E6 and E7 mRNA and protein expression were detected in cells treated by promoter-targeting siRNA. Futhermore, cellular growth, proliferation, apoptosis and senescence were detected. Thereafter, we investigated promoter DNA methylation and histone methylation status in cells treated with promoter-targeting siRNA. RESULTS: We found that E6/E7 mRNA and protein were simultaneously reduced, cell growth and proliferation were inhibited and cell death, especially senescence, was remarkably increased. Meanwhile, we also found a significantly increasing histone H3-Lys9 methylation on the promoter when E6/E7 gene expression was inhibited. INTERPRETATION: Our findings suggest that promoter-targeting siRNA effectively and simultaneously knocks down both extraneous HPV 16 E6 and E7 at the transcriptional level, and consequently inhibits proliferation and induces death in HPV 16-infected cells. This transcriptional repression is probably induced by histone modification rather than by alteration of DNA methylation.

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Section: E6 E7supporting

confidence: 78%

Gene silencing of HPV16 E6/E7 induced by promoter-targeting siRNA in SiHa cells

Hong

Lü

et al. 2009

Br J Cancer

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“…Since vertebrate genes are also characterized by extensive gene body methylation (Goll and Bestor, 2005;Klose and Bird, 2006), similar mechanisms could potentially apply for genes that are hypermethylated in cancer cells. In mammalian cells, a mechanistic link between siRNAs directed to promoters and induction of cytosine methylation has been convincingly demonstrated, which supports the basic existence of RdDMlike processes also in mammals (Morris et al, 2004;Castanotto et al, 2005). The existence of cancer cell or tumor-specific dsRNA is supported by a recent publication, which reports on natural antisense transcripts that are differentially expressed in various tumors (Klimov et al, 2006).…”

Section: Multiple Ways Of Recruiting Athda6?mentioning

confidence: 60%

“…However, similar mechanisms apparently exist in mammalian cells, suggesting the presence of functionally related components also in mammals. Short dsRNAs or siRNAs directed against promoters in mammalian cells cause transcriptional repression, but this is not always associated with DNA methylation (Morris et al, 2004;Castanotto et al, 2005;Ting et al, 2005;Weinberg et al, 2006). Although RNAi-mediated heterochromatin formation and RdDM have similar genetic requirements in Arabidopsis, suggesting a considerable overlap between these pathways, they can result in qualitatively distinct repressive chromatin at their targets (see below).…”

Section: Athda6: An Intimate Relationship With Transcriptional Rna Simentioning

confidence: 99%

Arabidopsis histone deacetylase 6: a green link to RNA silencing

et al. 2007

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Epigenetic reprogramming is at the base of cancer initiation and progression. Generally, genome-wide reduction in cytosine methylation contrasts with the hypermethylation of control regions of functionally wellestablished tumor suppressor genes and many other genes whose role in cancer biology is not yet clear. While insight into mechanisms that induce aberrant cytosine methylation in cancer cells is just beginning to emerge, the initiating signals for analogous promoter methylation in plants are well documented. In Arabidopsis, the silencing of promoters requires components of the RNA interference machinery and promoter double-stranded RNA (dsRNA) to induce a repressive chromatin state that is characterized by cytosine methylation and histone deacetylation catalysed by the RPD3-type histone deacetylase AtHDA6. Similar mechanisms have been shown to occur in fission yeast and mammals. This review focuses on the connections between cytosine methylation, dsRNA and AtHDA6-controlled histone deacetylation during promoter silencing in Arabidopsis and discusses potential mechanistic similarities of these silencing events in cancer and plant cells.

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“…TGS can be induced using siRNAs, antisense oligonucleotides [109], expressed antisense RNAs [110] or short hairpin RNAs [111,112]. And there are also examples of the successful implementation of this approach in vivo by local delivery in muscle [113], retina [112] and xenograft models [114].…”

Section: Gene-specific Epigenetic Therapymentioning

confidence: 99%

Epigenetics and ncRNAs in Brain Function and Disease: Mechanisms and Prospects for Therapy

2013

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The most fundamental roles of non-coding RNAs (ncRNAs) and epigenetic mechanisms are the guidance of cellular differentiation in development and the regulation of gene expression in adult tissues. In brain, both ncRNAs and the various epigenetic gene regulatory mechanisms play a fundamental role in neurogenesis and normal neuronal function. Thus, epigenetic chromatin remodelling can render coding sites transcriptionally inactive by DNA methylation, histone modifications or antisense RNA interactions. On the other hand, microRNAs (miRNAs) are ncRNA molecules that can regulate the expression of hundreds of genes post-transcriptionally, typically recognising binding sites in the 3′ untranslated region (UTR) of mRNA transcripts. Furthermore, there are a myriad of interactions in the interface of miRNAs and epigenetics. For example, epigenetic mechanisms can silence miRNA coding sites, and miRNAs can be the effectors of transcriptional gene silencing, targeting complementary promoters or silencing the expression of epigenetic modifier genes like MECP2 and EZH2 leading to global changes in the epigenome. Alterations in this regulatory machinery play a key role in the pathology of complex disorders including cancer and neurological diseases. For example, miRNA genes are frequently inactivated by epimutations in gliomas. Here we describe the interactions between epigenetic and ncRNA regulatory systems and discuss therapeutic potential, with an emphasis on tumors, cognitive disorders and neurodegenerative diseases.

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Short hairpin RNA-directed cytosine (CpG) methylation of the RASSF1A gene promoter in HeLa cells

Cited by 137 publications

References 18 publications

Gene silencing of HPV16 E6/E7 induced by promoter-targeting siRNA in SiHa cells

Gene silencing of HPV16 E6/E7 induced by promoter-targeting siRNA in SiHa cells

Arabidopsis histone deacetylase 6: a green link to RNA silencing

Epigenetics and ncRNAs in Brain Function and Disease: Mechanisms and Prospects for Therapy

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