DNA methylation inhibitor causes cell growth retardation and gene expression changes in feline lymphoma cells

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.

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DNA Methylation Inhibitor Regulates Guanylyl Cyclase/Natriuretic Peptide Receptor‐A Gene Expression in Mesangial Cells

The FASEB Journal ◽

10.1096/fasebj.29.1_supplement.880.25 ◽

2015 ◽

Vol 29 (S1) ◽

Author(s):

Prerna Kumar ◽

Kailash Pandey

Keyword(s):

Gene Expression ◽

Dna Methylation ◽

Natriuretic Peptide ◽

Guanylyl Cyclase ◽

Mesangial Cells ◽

Natriuretic Peptide Receptor ◽

Peptide Receptor ◽

Methylation Inhibitor ◽

Natriuretic Peptide Receptor A ◽

Dna Methylation Inhibitor

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Epigenetic, transcriptional and phenotypic responses in two generations of Daphnia magna exposed to the DNA methylation inhibitor 5-azacytidine

Environmental Epigenetics ◽

10.1093/eep/dvz016 ◽

2019 ◽

Vol 5 (3) ◽

Cited By ~ 7

Author(s):

Leif Christopher Lindeman ◽

Jens Thaulow ◽

You Song ◽

Jorke H Kamstra ◽

Li Xie ◽

...

Keyword(s):

Gene Expression ◽

Dna Methylation ◽

Histone Modifications ◽

Environmental Stressors ◽

Global Dna Methylation ◽

Two Generations ◽

Methylation Inhibitor ◽

Dna Methylation Inhibitor ◽

Offspring Generation ◽

Phenotypic Responses

Abstract The water flea Daphnia magna is a keystone species in freshwater ecosystems and has been widely used as a model organism in environmental ecotoxicology. This aquatic crustacean is sensitive to environmental stressors and displays considerable plasticity in adapting to changing environmental conditions. Part of this plasticity may be due to epigenetic regulation of gene expression, including changes to DNA methylation and histone modifications. Because of the generally hypomethylated genome of this species, we hypothesized that the histone code may have an essential role in the epigenetic control and that histone modifications might be an early marker for stress. This study aims to characterize the epigenetic, transcriptional and phenotypic responses and their causal linkages in directly exposed adult (F0) Daphnia and peritoneal exposed neonates (F1) after a chronic (7-day) exposure to a sublethal concentration (10 mg/l) of 5-azacytidine, a well-studied vertebrate DNA methylation inhibitor. Exposure of the F0 generation significantly reduced the cumulative fecundity, accompanied with differential expression of genes in the one-carbon-cycle metabolic pathway. In the epigenome of the F0 generation, a decrease in global DNA methylation, but no significant changes on H3K4me3 or H3K27me3, were observed. In the F1 offspring generation, changes in gene expression, a significant reduction in global DNA methylation and changes in histone modifications were identified. The results indicate that exposure during adulthood may result in more pronounced effects on early development in the offspring generation, though interpretation of the data should be carefully done since both the exposure regime and developmental period is different in the two generations examined. The obtained results improve our understanding of crustacean epigenetics and the tools developed may promote use of epigenetic markers in hazard assessment of environmental stressors.

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