Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Jul 5:409:51-8.
doi: 10.1016/j.mce.2015.03.023. Epub 2015 Apr 2.

Selective inhibition of prostaglandin E2 receptors EP2 and EP4 modulates DNA methylation and histone modification machinery proteins in human endometriotic cells

Joe A Arosh  1 , JeHoon Lee  2 , Anna Starzinski-Powitz  3 , Sakhila K Banu  2
Affiliations

Selective inhibition of prostaglandin E2 receptors EP2 and EP4 modulates DNA methylation and histone modification machinery proteins in human endometriotic cells

Joe A Arosh et al. Mol Cell Endocrinol. .

Abstract

Endometriosis is an inflammatory gynecological disease of reproductive-age women. The prevalence of endometriosis is 5-10% in reproductive-age women. Modern medical treatments are directed to inhibit the action of estrogen in endometriotic cells. However, hormonal therapies targeting estrogen can be prescribed only for a short time because of their undesirable side effects. Recent studies from our laboratory, using human endometriotic epithelial cell line 12Z and stromal cell line 22B derived from red lesion, discovered that selective inhibition of prostaglandin E2 (PGE2) receptors EP2 and EP4 inhibits adhesion, invasion, growth, and survival of 12Z and 22B cells by modulating integrins, MMPs and TIMPs, cell cycle, survival, and intrinsic apoptotic pathways, suggesting multiple epigenetic mechanisms. The novel findings of the present study indicate that selective pharmacological inhibition of EP2 and EP4: (i) decreases expression of DNMT3a, DNMT3b, H3K9me3, H3K27me3, SUV39H1, HP1a, H3K27, EZH2, JMJD2a, HDAC1, HDAC3, MeCP2, CoREST and Sin3A; (ii) increases expression of H3K4me3, H3H9ac, H3K27ac; and (iii) does not modulate the expression of DNMT1, hSET1, LSD1, MBD1, p300, HDAC2, and JMJD3 epigenetic machinery proteins in an epithelial and stromal cell specific manner. In this study, we report for the first time that inhibition of PGE2-EP2/EP4 signaling modulates DNA methylation, H3 histone methylation and acetylation, and epigenetic memory machinery proteins in human endometriotic epithelial cells and stromal cells. Thus, targeting EP2 and EP4 receptors may emerge as long-term nonsteroidal therapy for treatment of active endometriotic lesions in women.

Keywords: DNA methylation; EP2 and EP4; Endometriosis; Epigenetics; Histone modification; PGE(2).

PubMed Disclaimer

Figures

Figure 1:
Figure 1:. Effects of inhibition of EP2 and EP4 receptors on expression of DNA methyltransferases (DNMTs) in human endometriotic epithelial cells 12Z and stromal cells 22B.
(A1-A2) Western blot analysis and densitometry of (B1) DNMT1, (B2) DNMT3a, and (B3) DNMT3b proteins in 12Z and 22B cells. β-actin protein was measured as an internal control. The 12Z and 22B cells were treated with EP2 and EP4 inhibitors (EP-I) EP2 (AH6809–75 μM) and EP4 (AH23848- μM) for 24h. The experiments were performed as we described in the “Materials and Methods”. *-Control vs. EP2-I/EP4-I, P<0.05. Numerical data are expressed as Mean ± SEM of three (n=3) experiments.
Figure 2:
Figure 2:. Effects of inhibition of EP2 and EP4 receptors on H3K4, H3K9, and H3K27 methylation and acetylation in human endometriotic epithelial cells 12Z and stromal cells 22B.
(A1-A2) Western blot analysis and densitometry of proteins (B1) H3K4me3, (C1) H3K9ac, (C2) H3K9me3, (C3) H3K9me3: H3K9ac ratio, (D1) H3K27ac, (D2) H3K27me3, and (D3) H3K9me3: H3K9ac ratio in 12Z and 22B cells. β-actin protein was measured as an internal control. The 12Z and 22B cells were treated with EP2 and EP4 inhibitors (EP-I) EP2 (AH6809–75 μM) and EP4 (AH23848- μM) for 24h. The experiments were performed as we described in the “Materials and Methods”. *-Control vs. EP2-I/EP4-I, P<0.05. Numerical data are expressed as Mean ± SEM of three (n=3) experiments.
Figure 3:
Figure 3:. Effects of inhibition of EP2 and EP4 receptors on H3 histone methylation and demethylation machinery proteins in human endometriotic epithelial cells 12Z and stromal cells 22B.
(A1-A2) Western blot analysis and densitometry of proteins (B1) hSET1, (B2) HP1a, (B3) SUV39H1, (B4) EZH2, (B5) LSD1, (B6) JMJD2A, and (B7) JMJD3 in 12Z and 22B cells. β-actin protein was measured as an internal control. The 12Z and 22B cells were treated with EP2 and EP4 inhibitors (EP-I) EP2 (AH6809–75 μM) and EP4 (AH23848- μM) for 24h. The experiments were performed as we described in the “Materials and Methods”. *-Control vs. EP2-I/EP4-I, P<0.05. Numerical data are expressed as Mean ± SEM of three (n=3) experiments.
Figure 4:
Figure 4:. Effects of inhibition of EP2 and EP4 receptors on H3 histone acetylation and deacetylation machinery proteins in human endometriotic epithelial cells 12Z and stromal cells 22B.
(A1-A2) Western blot analysis and densitometry of proteins (B1) HDAC1, (B2) HDAC2, (B3) HDAC3, and (B4) p300 in endometriotic epithelial cells 12Z and stromal cells 22B. β-actin protein was measured as an internal control. The 12Z and 22B cells were treated with EP2 and EP4 inhibitors (EP-I) EP2 (AH6809–75 μM) and EP4 (AH23848- μM) for 24h. The experiments were performed as we described in the “Materials and Methods”. *-Control vs. EP2-I/EP4-I, P<0.05. Numerical data are expressed as Mean ± SEM of three (n=3) experiments.
Figure 5:
Figure 5:. Effects of inhibition of EP2 and EP4 receptors on transcriptional activation and suppression machinery proteins in human endometriotic epithelial cells 12Z and stromal cells 22B.
(A1-A2) Western blot analysis and densitometry of proteins (B1) Pol II, (B2) MeCP2, (B3) MBD1, (B4) CoREST, and (B5) Sin3A in endometriotic epithelial cells 12Z and stromal cells 22B. β-actin protein was measured as an internal control. The 12Z and 22B cells were treated with EP2 and EP4 inhibitors (EP-I) EP2 (AH6809–75 μM) and EP4 (AH23848- μM) for 24h. The experiments were performed as we described in the “Materials and Methods”. *-Control vs. EP2-I/EP4-I, P<0.05. Numerical data are expressed as Mean ± SEM of three (n=3) experiments.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Bulun SE (2009) Endometriosis. N Engl J Med 360, 268–79. - PubMed
    1. Giudice LC and Kao LC (2004) Endometriosis. Lancet 364, 1789–99. - PubMed
    1. Guo SW and Olive DL (2007) Two unsuccessful clinical trials on endometriosis and a few lessons learned. Gynecol Obstet Invest 64, 24–35. - PubMed
    1. Kyama CM, Mihalyi A, Simsa P, Mwenda JM, Tomassetti C, Meuleman C and D’Hooghe TM (2008) Non-steroidal targets in the diagnosis and treatment of endometriosis. Curr Med Chem 15, 1006–17. - PubMed
    1. Wu MH, Shoji Y, Chuang PC and Tsai SJ (2007) Endometriosis: disease pathophysiology and the role of prostaglandins. Expert Rev Mol Med 9, 1–20. - PubMed

Publication types

MeSH terms