scholarly journals SPOP mutation induces DNA methylation via stabilizing GLP/G9a

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jianong Zhang ◽  
Kun Gao ◽  
Hongyan Xie ◽  
Dejie Wang ◽  
Pingzhao Zhang ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Tumor Suppressor Genes ◽  
Dna Hypermethylation ◽  
Genome Wide ◽  
Anti Cancer ◽  
Methylome Analysis ◽  
Methylation Inhibitor ◽  
Dna Methylation Inhibitor ◽  
Underlying Mechanisms

AbstractMutations in SPOP E3 ligase gene are reportedly associated with genome-wide DNA hypermethylation in prostate cancer (PCa) although the underlying mechanisms remain elusive. Here, we demonstrate that SPOP binds and promotes polyubiquitination and degradation of histone methyltransferase and DNMT interactor GLP. SPOP mutation induces stabilization of GLP and its partner protein G9a and aberrant upregulation of global DNA hypermethylation in cultured PCa cells and primary PCa specimens. Genome-wide DNA methylome analysis shows that a subset of tumor suppressor genes (TSGs) including FOXO3, GATA5, and NDRG1, are hypermethylated and downregulated in SPOP-mutated PCa cells. DNA methylation inhibitor 5-azacytidine effectively reverses expression of the TSGs examined, inhibits SPOP-mutated PCa cell growth in vitro and in mice, and enhances docetaxel anti-cancer efficacy. Our findings reveal the GLP/G9a-DNMT module as a mediator of DNA hypermethylation in SPOP-mutated PCa. They suggest that SPOP mutation could be a biomarker for effective treatment of PCa with DNA methylation inhibitor alone or in combination with taxane chemotherapeutics.

167 EFFECT OF DNA METHYLATION INHIBITOR ON HETEROCHROMATIN IN BOVINE EMBRYOS DERIVED FROM HEAT-SHOCKED OOCYTES

2016 ◽  
Vol 28 (2) ◽  
pp. 213
Author(s):  
T. D. Araujo ◽  
J. Jasmin ◽  
C. C. R. Quintao ◽  
E. D. Souza ◽  
J. H. M. Viana ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Heat Shock ◽  
In Vitro Maturation ◽  
Methylation Status ◽  
Control Groups ◽  
Methylation Inhibitor ◽  
Response To Stress ◽  
Dna Methylation Inhibitor ◽  
And Control

The cell response to stress involves epigenetic modifications in order to regulate the gene expression, which is dependent of chromatin structure and DNA methylation status. On the other hand, changes on DNA methylation can have an effect on chromatin organisation (Espada and Esteller 2010 Semin. Cell. Dev. Biol. 21, 238–246). In this study we evaluated the effect of 5-aza-2′-deoxycytidine (5-aza; Sigma, St. Louis, MO, USA), a DNA methylation inhibitor, on heterochromatin 1 β formation of bovine pre-implantation embryos derived from oocytes that did or did not undergo heat shock during in vitro maturation (IVM). Oocytes were IVM under 38.5°C for 24 h (non-heat-shock: NHS group) or under 41.5°C for 12 h followed by 38.5°C for 12 h (heat-shock: HS group). Oocytes were IVF for 20 h and the denuded presumptive zygotes from NHS or HS groups were cultured with 0 (nontreated controls) or 10 nM of 5-aza for 24 h or 48 h in CR2aa plus 2.5% FCS at 38.5°C with 5% CO2, 5% O2 and 90% N2. Embryos with 4–7 cells at 44 h post-insemination (hpi) and embryos with 8–16 cells at 68 hpi were fixed in 4% paraformaldehyde and stained with anti-mouse HP1β first antibody, then immunofluorescence was evaluated by confocal microscopy (Leica TCS SP5II) and images were processed by ImageJ 1.49 (NIH, Bethesda, MD, USA). Fluorescence of nuclei and of background area (fluorescence/unit area) were measured and then the corrected relative fluorescence per nucleus was calculated. We analysed 129 and 149 nuclei at 44 hpi from 29 and 34 embryos as well as 268 and 182 nuclei at 68 hpi from 37 and 22 embryos of the NHS and HS groups, respectively, obtained from 3 replicates. Data underwent log-transformation and was analysed by ANOVA, and means compared by Student-Newman-Keuls. Embryos with 8–16 cells derived from NHS oocytes and treated with 5-aza for 24 h or for 48 h had nuclei with lower HP1 fluorescence than their respective NHS (nontreated) control (P < 0.01). In contrast, 8–16-cells embryos derived from HS and treated with 5-aza displayed nuclei with the same HP1 fluorescence of their respective HS control (P > 0.05). When embryos derived from HS and NHS (nontreated) control groups were compared, higher HP1 fluorescence was found in those with 4–7 cells of HS group (P < 0.05); however, embryos with 8–16 cells displayed similar HP1 fluorescence between both NHS and HS control groups (P > 0.05). There was no difference on HP1 fluorescence between nuclei of embryos with 4–7 cells treated with 5-aza for 24 h and control (nontreated) in both HS and NHS groups. These data suggest that embryos derived from heat-shocked oocytes can accumulate more heterochromatin at earlier stages than those from non-heat-shocked oocytes and that the effect of DNA methylation inhibition by 5-aza on embryo heterochromatin can vary accordingly to the exposure of the oocyte to heat shock during in vitro maturation. Financial support from CNPq, Fapemig, and CAPES is acknowledged.

scholarly journals Enhancement of In vitro and In vivo Tumor Cell Radiosensitivity by the DNA Methylation Inhibitor Zebularine

2005 ◽  
Vol 11 (12) ◽  
pp. 4571-4579 ◽  
Author(s):  
Hideaki Dote ◽  
David Cerna ◽  
William E. Burgan ◽  
Donna J. Carter ◽  
Michael A. Cerra ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Tumor Cell ◽  
Methylation Inhibitor ◽  
Dna Methylation Inhibitor

scholarly journals Genome-wide epigenetic analysis delineates a biologically distinct immature acute leukemia with myeloid/T-lymphoid features

Blood ◽  
2009 ◽  
Vol 113 (12) ◽  
pp. 2795-2804 ◽  
Author(s):  
Maria E. Figueroa ◽  
Bas J. Wouters ◽  
Lucy Skrabanek ◽  
Jacob Glass ◽  
Yushan Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Cpg Islands ◽  
Specific Gene ◽  
Molecular Heterogeneity ◽  
Dna Hypermethylation ◽  
Genome Wide

Abstract Acute myeloid leukemia is a heterogeneous disease from the molecular and biologic standpoints, and even patients with a specific gene expression profile may present clinical and molecular heterogeneity. We studied the epigenetic profiles of a cohort of patients who shared a common gene expression profile but differed in that only half of them harbored mutations of the CEBPA locus, whereas the rest presented with silencing of this gene and coexpression of certain T-cell markers. DNA methylation studies revealed that these 2 groups of patients could be readily segregated in an unsupervised fashion based on their DNA methylation profiles alone. Furthermore, CEBPA silencing was associated with the presence of an aberrant DNA hypermethylation signature, which was not present in the CEBPA mutant group. This aberrant hypermethylation occurred more frequently at sites within CpG islands. CEBPA-silenced leukemias also displayed marked hypermethylation compared with normal CD34+ hematopoietic cells, whereas CEBPA mutant cases showed only mild changes in DNA methylation compared with these normal progenitors. Biologically, CEBPA-silenced leukemias presented with a decreased response to myeloid growth factors in vitro.

scholarly journals Analysis of Gene Expression Patterns of Epigenetic Enzymes Dnmt3a, Tet1 and Ogt in Murine Chondrogenic Models

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2678
Author(s):  
Judit Vágó ◽  
Katalin Kiss ◽  
Edina Karanyicz ◽  
Roland Takács ◽  
Csaba Matta ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
In Situ Hybridization ◽  
Marker Genes ◽  
Specific Gene ◽  
Cartilage Formation ◽  
Methylation Inhibitor ◽  
Dna Methylation Inhibitor

We investigated the gene expression pattern of selected enzymes involved in DNA methylation and the effects of the DNA methylation inhibitor 5-azacytidine during in vitro and in vivo cartilage formation. Based on the data of a PCR array performed on chondrifying BMP2-overexpressing C3H10T1/2 cells, the relative expressions of Tet1 (tet methylcytosine dioxygenase 1), Dnmt3a (DNA methyltransferase 3), and Ogt (O-linked N-acetylglucosamine transferase) were further examined with RT-qPCR in murine cell line-based and primary chondrifying micromass cultures. We found very strong but gradually decreasing expression of Tet1 throughout the entire course of in vitro cartilage differentiation along with strong signals in the cartilaginous embryonic skeleton using specific RNA probes for in situ hybridization on frozen sections of 15-day-old mouse embryos. Dnmt3a and Ogt expressions did not show significant changes with RT-qPCR and gave weak in situ hybridization signals. The DNA methylation inhibitor 5-azacytidine reduced cartilage-specific gene expression and cartilage formation when applied during the early stages of chondrogenesis. In contrast, it had a stimulatory effect when added to differentiated chondrocytes, and quantitative methylation-specific PCR proved that the DNA methylation pattern of key chondrogenic marker genes was altered by the treatment. Our results indicate that the DNA demethylation inducing Tet1 plays a significant role during chondrogenesis, and inhibition of DNA methylation exerts distinct effects in different phases of in vitro cartilage formation.

scholarly journals The Methylation Inhibitor 5-Aza-2′-Deoxycytidine Induces Genome-Wide Hypomethylation in Rice

2021 ◽  
Author(s):  
Shuo Liu ◽  
Yu Bao ◽  
Hui Deng ◽  
Guanqing Liu ◽  
Yangshuo Han ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Genome Stability ◽  
Epigenetic Modification ◽  
Homozygous Mutation ◽  
Global Methylation ◽  
Genome Wide ◽  
Methylation Inhibitor ◽  
Dna Methylation Inhibitor ◽  
Genomic Regions ◽  
Genome Scale

Abstract DNA methylation is a conserved epigenetic modification which is vital for regulating gene expression and maintaining genome stability in both mammals and plants. Homozygous mutation of rice methyltransferase 1 (met1) gene can cause host death in rice, making it difficult to obtain plant material needed for hypomethylation research. To circumvent this challenge, the methylation inhibitor, 5-Aza-2′-deoxycytidine (AzaD), is used as a cytosine nucleoside analogue to reduce genome wide hypomethylation and is widely used in hypomethylation research. However, how AzaD affects plant methylation profiles at the genome scale is largely unknown. Here, we treated rice seedlings with AzaD and compared the AzaD treatment with osmet1-2 mutants, illustrating that there are similar CG hypomethylation and distribution throughout the whole genome. Along with global methylation loss class I transposable elements (TEs) which are farther from genes compared with class II TEs, were more significantly activated, and the RNA-directed DNA Methylation (RdDM) pathway was activated in specific genomic regions to compensate for severe CG loss. Overall, our results suggest that AzaD is an effective DNA methylation inhibitor that can influence genome wide methylation and cause a series of epigenetic variations.

scholarly journals Integrated Methylome and Transcriptome Analyses Reveal the Molecular Mechanism by Which DNA Methylation Regulates Kenaf Flowering

2021 ◽  
Vol 12 ◽  
Author(s):  
Zengqiang Li ◽  
Meiqiong Tang ◽  
Dengjie Luo ◽  
Muhammad Haneef Kashif ◽  
Shan Cao ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Underlying Mechanism ◽  
Differentially Expressed ◽  
Genome Wide ◽  
Differentially Methylated Genes ◽  
Methylation Inhibitor ◽  
Dna Methylation Inhibitor ◽  
Transcriptome Analyses ◽  
Flowering Regulation ◽  
Dna Methylation Analysis

DNA methylation regulates key biological processes in plants. In this study, kenaf seedlings were pretreated with the DNA methylation inhibitor 5-azacytidine (5-azaC) (at concentrations of 0, 100, 200, 400, and 600 μM), and the results showed that pretreatment with 200 μM 5-azaC promoted flowering most effectively. To elucidate the underlying mechanism, phytohormone, adenosine triphosphate (ATP), and starch contents were determined, and genome-wide DNA methylation and transcriptome analyses were performed on anthers pretreated with 200 μM 5-azaC (5-azaC200) or with no 5-azaC (control conditions; 5-azaC0). Biochemical analysis revealed that 5-azaC pretreatment significantly reduced indoleacetic acid (IAA) and gibberellic acid (GA) contents and significantly increased abscisic acid (ABA) and ATP contents. The starch contents significantly increased in response to 200 and 600 μM 5-azaC. Further genome-wide DNA methylation analysis revealed 451 differentially methylated genes (DMGs) with 209 up- and 242 downregulated genes. Transcriptome analysis showed 3,986 differentially expressed genes (DEGs), with 2,171 up- and 1,815 downregulated genes. Integrated genome-wide DNA methylation and transcriptome analyses revealed 72 genes that were both differentially methylated and differentially expressed. These genes, which included ARFs, PP2C, starch synthase, FLC, PIF1, AGL80, and WRKY32, are involved mainly in plant hormone signal transduction, starch and sucrose metabolism, and flowering regulation and may be involved in early flowering. This study serves as a reference and theoretical basis for kenaf production and provides insights into the effects of DNA methylation on plant growth and development.

scholarly journals Combination of Gefitinib and DNA Methylation Inhibitor Decitabine Exerts Synergistic Anti-Cancer Activity in Colon Cancer Cells

PLoS ONE ◽  
2014 ◽  
Vol 9 (5) ◽  
pp. e97719 ◽  
Author(s):  
Yun-feng Lou ◽  
Zheng-zhi Zou ◽  
Pin-jia Chen ◽  
Guo-bin Huang ◽  
Bin Li ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Colon Cancer ◽  
Cancer Cells ◽  
Colon Cancer Cells ◽  
Anti Cancer ◽  
Methylation Inhibitor ◽  
Dna Methylation Inhibitor ◽  
Cancer Activity

Novel Findings of Anti-cancer Property of Propofol

2018 ◽  
Vol 18 (2) ◽  
pp. 156-165 ◽  
Author(s):  
Jiaqiang Wang ◽  
Chien-shan Cheng ◽  
Yan Lu ◽  
Xiaowei Ding ◽  
Minmin Zhu ◽  
...  
Keyword(s):  
General Anesthesia ◽  
Cancer Progression ◽  
Molecular Mechanisms ◽  
Intravenous Anesthetic ◽  
The Past ◽  
Anti Cancer ◽  
Underlying Mechanisms ◽  
Recent Attention

Background: Propofol, a widely used intravenous anesthetic agent, is traditionally applied for sedation and general anesthesia. Explanation: Recent attention has been drawn to explore the effect and mechanisms of propofol against cancer progression in vitro and in vivo. Specifically, the proliferation-inhibiting and apoptosis-inducing properties of propofol in cancer have been studied. However, the underlying mechanisms remain unclear. Conclusion: This review focused on the findings within the past ten years and aimed to provide a general overview of propofol's malignance-modulating properties and the potential molecular mechanisms.

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