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Regulation of DNA methyltransferase 3a Expression by Prostaglandin E2 in Human Fibroblasts
Author(s) -
Seutter Sara,
Mercando Jason,
Sen Neil,
Penev Boris,
Auker Bennet,
Shamseddin Seyed M.,
Green Kevan,
Borghaei Ruth
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.03050
Subject(s) - dnmt1 , dna methylation , context (archaeology) , methyltransferase , receptor , dna methyltransferase , methylation , gene expression , epigenetics , biology , cancer research , inflammation , microbiology and biotechnology , signal transduction , gene , immunology , genetics , paleontology
Periodontitis is common chronic inflammatory condition that not only results in loss of teeth, but contributes to a variety of systemic conditions, including cancer. Previously we showed that exposure of human gingival fibroblasts (HGF) derived from patients with periodontitis to PGE2 decreases expression of DNMT1, DNMT3a and TET1, but increases global levels of methylated and hydroxymethylated DNA, acting predominantly through the EP4 receptor. The current study compares the effects of PGE2 on expression of these genes in non‐inflamed fibroblasts and aims to further elucidate the signaling pathways involved. Methods Real time PCR and ELISA are used to measure expression of DNMT1, DNMT3a, TET1 and EP receptors 1–4 in untreated HGF and IMR90 lung fibroblasts and those treated with PGE2. The effects of actinomycin D, as well as EP receptor agonists and antagonists and signaling pathway activators and inhibitors on mRNA expression of these genes will also be assessed by qPCR. Results Results so far show that PGE2 decreases expression of DNMT1 in IMR90 cells to an extent similar to that observed in HGF, but levels of DNMT3a are increased rather than decreased, and levels of TET1 are unaffected in IMR90 cells. Although all four EP receptors are expressed in both HGF and IMR90, the relative levels of EP4 expression are lower. Conclusion The ability of this inflammatory mediator to influence DNA methylation is consistent with the association of chronic inflammation with increased risk of cancer. However, the fact that the effects are cell and context dependent indicate that more information about the specific signaling pathways involved in regulating expression of these important enzymes is needed. Further studies will be aimed at elucidating the roles of the PGE2 receptors and their signaling pathways in mediating the cell‐specific effects.