scholarly journals DNA Methylation Patterns Correlate with the Expression of SCNN1A, SCNN1B, and SCNN1G (Epithelial Sodium Channel, ENaC) Genes

2021 ◽  
Vol 22 (7) ◽  
pp. 3754
Author(s):  
Silvia Pierandrei ◽  
Gessica Truglio ◽  
Fabrizio Ceci ◽  
Paola Del Porto ◽  
Sabina Maria Bruno ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Dna Methylation ◽  
Sodium Channel ◽  
Cell Lines ◽  
Transcriptional Control ◽  
Inverse Correlation ◽  
Epithelial Sodium Channel ◽  
Methylation Pattern ◽  
Methylation Patterns ◽  
Epithelial Sodium Channel Enac

The interplay between the cystic fibrosis transmembrane conductance regulator (CFTR) and the epithelial sodium channel (ENaC) in respiratory epithelia has a crucial role in the pathogenesis of cystic fibrosis (CF). The comprehension of the mechanisms of transcriptional regulation of ENaC genes is pivotal to better detail the pathogenic mechanism and the genotype–phenotype relationship in CF, as well as to realize therapeutic approaches based on the transcriptional downregulation of ENaC genes. Since we aimed to study the epigenetic transcriptional control of ENaC genes, an assessment of their expression and DNA methylation patterns in different human cell lines, nasal brushing samples, and leucocytes was performed. The mRNA expression of CFTR and ENaC subunits α, β and γ (respectively SCNN1A, SCNN1B, and SCNN1G genes) was studied by real time PCR. DNA methylation of 5′-flanking region of SCNN1A, SCNN1B, and SCNN1G genes was studied by HpaII/PCR. The levels of expression and DNA methylation of ENaC genes in the different cell lines, brushing samples, and leukocytes were very variable. The DNA regions studied of each ENaC gene showed different methylation patterns. A general inverse correlation between expression and DNA methylation was evidenced. Leukocytes showed very low expression of all the 3 ENaC genes corresponding to a DNA methylated pattern. The SCNN1A gene resulted to be the most expressed in some cell lines that, accordingly, showed a completely demethylated pattern. Coherently, a heavy and moderate methylated pattern of, respectively, SCNN1B and SCNN1G genes corresponded to low levels of expression. As exceptions, we found that dexamethasone treatment appeared to stimulate the expression of all the 3 ENaC genes, without an evident modulation of the DNA methylation pattern, and that in nasal brushing a considerable expression of all the 3 ENaC genes were found despite an apparent methylated pattern. At least part of the expression modulation of ENaC genes seems to depend on the DNA methylation patterns of specific DNA regions. This points to epigenetics as a controlling mechanism of ENaC function and as a possible therapeutic approach for CF.

Analysis of Methylation Status of ZAR1, GATA4, CDH22, SOX3, SLC16A5, PFN2, EHD3 and TBPL1 in Hematological Malignancies

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4631-4631 ◽  
Author(s):  
Yujin Kobayashi ◽  
Yoshihiro Hatta ◽  
Jin Takeuchi ◽  
Kyoko Fujiwara ◽  
Yui Shinojima ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Cell Line ◽  
Cell Lines ◽  
Hematological Malignancies ◽  
Methylation Pattern ◽  
Human Leukemia ◽  
Aberrant Methylation ◽  
Methylation Analysis ◽  
Clinical Specimens ◽  
Methylation Patterns

Abstract Abstract 4631 Background: DNA cytosine methylation in CpG dinucleotides is an important epigenetic event and critical for the control of gene expression, and appears to play a crucial role in tumorgenesis in hematologic malignancies. To identify novel tumor-specific differentially methylated regions in human leukemia/lymphoma, we performed methylation analysis of new target lesions in hematological malignancies. Methods: The aberrant methylation at 8 new candidate human homologous genomic regions (ZAR1, GATA4, CDH22, SOX3, SLC16A5, PFN2, EHD3 and TBPL1), where hypermethyloation status was identified through a mouse-skin cancer model study by the RLSG method, were analysed with quantitative DNA methylation analysis using the Sequenom MassARRAY system. 8 human leukemia/lymphoma cell lines (HL-60, KG-1, Jurkat, MOLT-4, NALM-6, K562, Ramos, and Raji), and bone marrow or peripheral blood samples from 20 patients with leukemia before treatment were obtained for samples (10 in AML, 4 in Ph- ALL, 2 in Ph+ ALL, 2 in CML-CP, 1 in CML-BC, and 1 in CLL). Normal lymphocyte cells from four healthy individuals were used as normal controls. Each data were assigned as hypermethylaion when the average methylation levels of the entire target regions were more than 50%. Results: In ZAR1, SLC16A5 and EHD3, hypermethylation level was seen in all leukemia/lymphoma cell lines. GATA4, CDH22 and SOX3 showed hypermethylaion in all tumor cell lines except for AML cell line KG-1. PFN2 showed hypermethylation in all cell lines except for KG-1, T-ALL cell line Jurkat and MOLT-4. No aberrant methylation among tumor and normal cells was observed in TBPL1. Interestingly, DNA methylation patterns of ZAR1, GATA4, CDH22, and SOX3 were different between the clinical specimens of AML and ALL. Hypermethylation of those genes were frequently observed in ALL samples but less in AML. The Mann-Whitney U-test showed that the differences were significant in all ZAR1, GATA4, CDH22, and SOX3 (p < 0.01, respectively). DNA samples from CML-CP patients showed no abnormal methylation patterns in all genes. DNA from T-CLL patient showed hypermethylation only in EHD3. No abnormal methylation pattern was observed in SLC16A5 and PFN2 in clinical specimens. None of the normal control samples showed aberrant methylation in any genes. Conclusion: We identified regions aberrantly methylated with high frequency among the new candidate genes in leukemia and lymphoma, and demonstrated a distinct methylation pattern between tumors and normal lymphocytes. Aberrant DNA methylation of ZAR1, GATA4, CDH22 and SOX3 may be associated to differentiation to lymphoid populations of leukemia. Disclosures: No relevant conflicts of interest to declare.

Examination of the dynamics of global DNA methylation pattern establishment during spermatogenesiss

2007 ◽  
Vol 30 (4) ◽  
pp. 90
Author(s):  
Kirsten Niles ◽  
Sophie La Salle ◽  
Christopher Oakes ◽  
Jacquetta Trasler
Keyword(s):  
Dna Methylation ◽  
Germ Cell ◽  
Germ Cells ◽  
Epigenetic Modification ◽  
Methylation Pattern ◽  
Chromosome 9 ◽  
Specific Gene ◽  
Global Methylation ◽  
Dna Methylation Pattern ◽  
Methylation Patterns

Background: DNA methylation is an epigenetic modification involved in gene expression, genome stability, and genomic imprinting. In the male, methylation patterns are initially erased in primordial germ cells (PGCs) as they enter the gonadal ridge; methylation patterns are then acquired on CpG dinucleotides during gametogenesis. Correct pattern establishment is essential for normal spermatogenesis. To date, the characterization and timing of methylation pattern acquisition in PGCs has been described using a limited number of specific gene loci. This study aimed to describe DNA methylation pattern establishment dynamics during male gametogenesis through global methylation profiling techniques in a mouse model. Methods: Using a chromosome based approach, primers were designed for 24 regions spanning chromosome 9; intergenic, non-repeat, non-CpG island sequences were chosen for study based on previous evidence that these types of sequences are targets for testis-specific methylation events. The percent methylation was determined in each region by quantitative analysis of DNA methylation using real-time PCR (qAMP). The germ cell-specific pattern was determined by comparing methylation between spermatozoa and liver. To examine methylation in developing germ cells, spermatogonia from 2 day- and 6 day-old Oct4-GFP (green fluorescent protein) mice were isolated using fluorescence activated cell sorting. Results: As compared to liver, four loci were hypomethylated and five loci were hypermethylated in spermatozoa, supporting previous results indicating a unique methylation pattern in male germ cells. Only one region was hypomethylated and no regions were hypermethylated in day 6 spermatogonia as compared to mature spermatozoa, signifying that the bulk of DNA methylation is established prior to type A spermatogonia. The methylation in day 2 spermatogonia, germ cells that are just commencing mitosis, revealed differences of 15-20% compared to day 6 spermatogonia at five regions indicating that the most crucial phase of DNA methylation acquisition occurs prenatally. Conclusion: Together, these studies provide further evidence that germ cell methylation patterns differ from those in somatic tissues and suggest that much of methylation at intergenic sites is acquired during prenatal germ cell development. (Supported by CIHR)

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