The estrogen metabolite 2-methoxyestradiol (2-ME2) is one of the most potent antiangiogenic and proapoptotic endogenous steroids. Herein, we investigate the effects of 2-ME2 on angiogenesis of cultured primary ovine uterine artery endothelial cells (UAECs) from nonpregnant follicular (F-UAECs), nonpregnant luteal (L-UAECs), and pregnant ewes (P-UAECs). Uterine artery endothelial cells were treated with vehicle control, 10−8 mol/L 17β-estradiol (17βE2), or 10−9 to 10−6 mol/L 2-ME2. Angiogenesis, apotosis, and cell morphology were assessed by capillary tube formation, flowcytometry, and immunohistochemistry. 17βE2 stimulated while 10−6 mol/L 2-ME2 inhibited capillary tube formation in F-UAECs (P < .05). The inhibitory effects of 2-ME2 on angiogenesis were minimal in L-UAECs and were absent in P-UAECs when compared to controls. 10−6 mol/L 2-ME2 increased apoptosis and inhibited microtubular structure equally in pregnant and nonpregnant UAECs when compared to control or 17βE2 treatments. Thus, 2-ME2 inhibit capillary tube formation in F-UAECs while L-UAECs and P-UAECs are relatively unresponsive to the inhibitory effects of 2ME2 indicating that the pregnancy phenotypic state of the UAECs may modulate the action of 2-ME2 on capillary angiogenesis.
JohnsonMLGrazul-BilskaATRedmerDAReynoldsLP. Effects of estradiol-17beta on expression of mRNA for seven angiogenic factors and their receptors in the endometrium of ovariectomized (OVX) ewes. Endocrine. 2006;30(3):333–342.
2.
PepeGJAlbrechtED. Regulation of functional differentiation of the placental villous syncytiotrophoblast by estrogen during primate pregnancy. Steroids. 1999;64(9):624–627.
3.
RubanyiGMFreayADKauserK. Vascular estrogen receptors and endothelium-derived nitric oxide production in the mouse aorta. Gender difference and effect of estrogen receptor gene disruption. J Clin Invest. 1997;99(10):2429–2437.
4.
AberdeenGWBaschatAAHarmanCR. Uterine and fetal blood flow indexes and fetal growth assessment after chronic estrogen suppression in the second half of baboon pregnancy. Am J Physiol Heart Circ Physiol. 2010;298(3):H881–H889.
5.
ChangKLuboZ. Review article: steroid hormones and uterine vascular adaptation to pregnancy. Reprod Sci. 2008;15(4):336–348.
6.
ZachariaLCGogosJAKarayiorgouM. Methoxyestradiols mediate the antimitogenic effects of 17beta-estradiol: direct evidence from catechol-O-methyltransferase-knockout mice. Circulation. 2003;108(24):2974–2978.
7.
DubeyRKJacksonEKGillespieDG. Cytochromes 1A1/1B1- and catechol-O-methyltransferase-derived metabolites mediate estradiol-induced antimitogenesis in human cardiac fibroblast. J Clin Endocrinol Metab. 2005;90(1):247–255.
8.
JobeSORamadossJKochJMJiangYZhengJMagnessRR. Estradiol-17beta and its cytochrome P450- and catechol-O-methyltransferase-derived metabolites stimulate proliferation in uterine artery endothelial cells: role of estrogen receptor-alpha versus estrogen receptor-beta. Hypertension. 2010;55(4):1005–1011.
9.
D'AmatoRJLinCMFlynnEFolkmanJHamelE. 2-Methoxyestradiol, an endogenous mammalian metabolite, inhibits tubulin polymerization by interacting at the colchicine site. Proc Natl Acad Sci U S A. 1994;91(9):3964–3968.
10.
KlauberNParangiSFlynnEHamelED'AmatoRJ. Inhibition of angiogenesis and breast cancer in mice by the microtubule inhibitors 2-methoxyestradiol and taxol. Cancer Res. 1997;57(1):81–86.
11.
YueTLWangXLoudenCS. 2-Methoxyestradiol, an endogenous estrogen metabolite, induces apoptosis in endothelial cells and inhibits angiogenesis: possible role for stress-activated protein kinase signaling pathway and Fas expression. Mol Pharmacol. 1997;51(6):951–962.
12.
BrownJWKeslerCTNearyTFishmanLM. Effects of androgens and estrogens and catechol and methoxy-estrogen derivatives on mitogen-activated protein kinase (ERK(1,2)) activity in SW-13 human adrenal carcinoma cells. Horm Metab Res. 2001;33(3):127–130.
13.
MabjeeshNJEscuinDLaValleeTM. 2ME2 inhibits tumor growth and angiogenesis by disrupting microtubules and dysregulating HIF. Cancer Cell. 2003;3(4):363–375.
14.
DubeyRKGillespieDGZachariaLC. Methoxyestradiols mediate the antimitogenic effects of estradiol on vascular smooth muscle cells via estrogen receptor-independent mechanisms. Biochem Biophys Res Commun. 2000;278(1):27–33.
15.
MannistoPTKaakkolaS. Catechol-O-methyltransferase (COMT): biochemistry, molecular biology, pharmacology, and clinical efficacy of the new selective COMT inhibitors. Pharmacol Rev. 1999;51(4):593–628.
16.
MerriamGRMacLuskyNJPicardMKNaftolinF. Comparative properties of the catechol estrogens, I: methylation by catechol-O-methyltransferase and binding to cytosol estrogen receptors. Steroids. 1980;36(1):1–11.
17.
BasuAHaldarS. 2-Methoxyestradiol mediated signaling network in pancreatic cancer. Front Biosci. 2009;14:2170–2180.
18.
KanasakiKPalmstenKSugimotoH. Deficiency in catechol-O-methyltransferase and 2-methoxyoestradiol is associated with pre-eclampsia. Nature. 2008;453(7198):1117–1121.
19.
FotsisTZhangYPepperMS. The endogenous oestrogen metabolite 2-methoxyoestradiol inhibits angiogenesis and suppresses tumour growth. Nature. 1994;368(6468):237–239.
20.
BirdIMSullivanJADiT. Pregnancy-dependent changes in cell signaling underlie changes in differential control of vasodilator production in uterine artery endothelial cells. Endocrinology. 2000;141(3):1107–1117.
21.
ChenDBBirdIMZhengJMagnessRR. Membrane estrogen receptor-dependent extracellular signal-regulated kinase pathway mediates acute activation of endothelial nitric oxide synthase by estrogen in uterine artery endothelial cells. Endocrinology. 2004;145(1):113–125.
22.
ZhengJWenYChenDBBirdIMMagnessRR. Angiotensin II elevates nitric oxide synthase 3 expression and nitric oxide production via a mitogen-activated protein kinase cascade in ovine fetoplacental artery endothelial cells. Biol Reprod. 2005;72(6):1421–1428.
23.
MartinSJReutelingspergerCPMcGahonAJ. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J Exp Med. 1995;182(5):1545–1556.
24.
SalihSMSalamaSAFadlAANagamaniMAl-HendyA. Expression and cyclic variations of catechol-O-methyl transferase in human endometrial stroma. Fertil Steril. 2008;90(3):789–797.
25.
RajakumarABrandonHMDaftaryANessRConradKP. Evidence for the functional activity of hypoxia-inducible transcription factors overexpressed in preeclamptic placentae. Placenta. 2004;25(10):763–769.
26.
GogosJAMorganMLuineV. Catechol-O-methyltransferase-deficient mice exhibit sexually dimorphic changes in catecholamine levels and behavior. Proc Natl Acad Sci U S A. 1998;95(17):9991–9996.
27.
ZachariaLCJacksonEKGillespieDGDubeyRK. Catecholamines abrogate antimitogenic effects of 2-hydroxyestradiol on human aortic vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 2001;21(11):1745–1750.
28.
BarneaERAvigdorS. Coordinated induction of estrogen hydroxylase and catechol-O-methyl transferase by xenobiotics in first trimester human placental explants. J Steroid Biochem. 1990;35(2):327–331.
