Objective: To identify and characterize estrogen receptors in human umbilical vascular tissues and in cultured cells derived from the human umbilical vein.
Methods: Human umbilical vein endothelial (HUVE) and human umbilical vein smooth muscle (HUVSM) cells were isolated. Immunohistochemical, radioligand binding, Western immunoblotting, and reverse transcription-polymerase chain reation (RT-PCR) methods were used to detect estrogen receptors in vascular tissues and in cells derived from the umbilical cord.
Results: Estrogen receptor protein was not detected in either umbilical vessel tissue or in isolated HUVE or HUVSM cells. Messenger RNAs for the classic estrogen receptor (α) and estrogen receptor β isoforms also were undetectable by RT-PCR.
Conclusion: These findings suggest that the effects of estradiol observed in this widely used vascular model are mediated by very low concentrations of receptors that evade standard methods of detection. Alternatively, this steroid may affect umbilical vascular cells through mechanisms that do not involve the classic genomic estrogen-receptor pathway.
1. Trudinger BJ, Giles WB, Cook CM, Bombardieri J, Collins L. Fetal umbilical artery flow velocity waveforms and placental resistance: Clinical significance. Br J Obstet Gynaecol1985;92:23-30.
2.
2. Laurin J, Marsal K, Persson PH, Lingman G. Ultrasound measurement of fetal blood flow in predicting fetal outcome. Br J Obstet Gynaecol1987;94:940-948.
3.
3. Divon MY. Umbilical artery Doppler velocimetry: Clinical utility in high-risk pregnancies. Am J Obstet Gynecol1996;174:10-14.
4.
4. Challis DE, Warren PS, Gill RW. The significance of high umbilical venous blood flow measurements in a high-risk population. J Ultrasound Med1995;14:907-912.
5.
5. Resnik R, Killam AP, Battaglia FC, Makowski EL, Meschia G. The stimulation of uterine blood flow by various estrogens. Endocrinology1974;94:1192-1196.
6.
6. Veille JC, Li P, Eisenach JC, Massmann AG, Figueroa JP. Effects of estrogen on nitric oxide biosynthesis and vasorelaxant activity in sheep uterine and renal arteries in vitro. Am J Obstet Gynecol1996;17:1043-1049.
7.
7. Farhat MY, Lavigne MC, Raniwell PW. The vascular protective effects of estrogen. FASEB J1996;10:615-624.
8.
8. Bonatz G, Lorcher E, Weisner D, Schulz V, Semm K. Value of pulsatile Doppler ultrasound in diagnosis of suspected placental insufficiency in comparison with other examination parameters. Zentralbl Gynakol1996;118:129-132.
9.
9. Waddell BJ, Pepe GJ, Albrecht ED. Progesterone and 20 alpha-hydroxypregn-4-en-3-one (20 alpha-OHP) in the pregnant baboon: Selective placental secretion of 20 alpha-OHP into the fetal compartment. Biol Reprod1996;55:854-859.
10.
10. Magness RR, Rosenfeld CR, Carr BR. Protein kinase C in uterine and systemic arteries during ovarian cycle and pregnancy. Am J Physiol1991;260:E464-E470.
11.
11. Stice SL, Ford SP, Rosazza JP, Van Orden DE. Interaction of 4-hydroxylated estradiol and potential-sensitive Ca2+ channels in altering uterine blood flow during the estrous cycle and early pregnancy in gilts. Biol Reprod1987;36:369-375.
12.
12. Bush TL, Barrett-Connor E, Cowan LD, et al. Cardiovascular mortality and noncontraceptive use of estrogen in women: Results from the Lipid Research Clinics Program Follow-Up Study. Circulation1987;75:1102-1109.
13.
13. Gangar KF, Vyas S, Whitehead M, Crook D, Meire H, Campbell S. Pulsatility index in internal carotid artery in relation to transdermal oestradiol and time since menopause. Lancet1991;338:839-842.
14.
14. Pines A, Fisman EZ, Levo Y, et al. The effects of hormone replacement therapy in normal postmenopausal women: Measurements of Doppler-derived parameters of aortic flow. Am J Obstet Gynecol1991;164:806-812.
15.
15. Lieberman EH, Gerhard MD, Uehata A, et al. Estrogen improves endothelium-dependent, flow-mediated vasodilation in post-menopausal women. Ann Intern Med1994;121:936-941.
16.
16. Gilligan DM, Badar DM, Panza JA, Quyyumi AA, Cannon RO III. Effects of estrogen replacement therapy on peripheral vasomotor function in postmenopausal women. Am J Cardiol1995;75:264-268.
17.
17. Cheng LP, Kuwahara M, Jacobsson J, Foegh ML. Inhibition of myointimal hyperplasia and macrophage infiltration by estradiol in aorta allografts. Transplantation1991;52:967-972.
18.
18. Karas RH, Patterson BL, Mendelsohn ME. Human vascular smooth muscle cells contain functional estrogen receptor. Circulation1994;89:1943-1950.
19.
19. Mugge A, Riedel M, Barton M, Kuhn M, Lichtlen PR. Endothelium independent relaxation of human coronary arteries by 17 beta-oestradiol in vitro. Cardiovasc Res1993;27:1939-1942.
20.
20. Perrot-Applanat M, Groyer-Picard MT, Garcia E, Lorenzo F, Milgrom E. Immunocytochemical demonstration of estrogen and progesterone receptors in muscle cells of uterine arteries in rabbits and humans. Endocrinology1988;123:1511-1519.
21.
21. Losordo DW, Kearney M, Kim EA, Jekanowski J, Isner JM. Variable expression of the estrogen receptor in normal and atherosclerotic coronary arteries of premenopausal women. Circulation1994;89:1501-1510.
22.
22. Pertscuk L. ERICA and PgRICA in the umbilical cord, endothelial cell culture, and kidney. In: Immunocytochemistry for steroid receptors. Boca Raton, Florida: CRC Press, 1990:181-190.
23.
23. Hayashi T, Yamada K, Esaki T, et al. Estrogen increases endothelial nitric oxide by a receptor-mediated system. Biochem Biophys Res Commun1995;214:847-855.
24.
24. Venkov CD, Rankin AB, Vaughan DE. Identification of authentic estrogen receptor in cultured endothelial cells. A potential mechanism for steroid hormone regulation of endothelial function. Circulation1996;94:727-733.
25.
25. Caulin-Glaser T, Watson CA, Pardi R, Bender JR. Effects of 17β-estradiol on cytokine-induced endothelial cell adhesion molecule expression. J. Clin Invest1996;98:36-42.
26.
26. Fox SB, Khong TY. Lack of innervation of human umbilical cord. An immunohistological and histochemical study. Placenta1990;11:59-62.
27.
27. Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest1973;52:2745-2756.
28.
28. Gimbrone Ma, Cotran RS. Human vascular smooth muscle in culture. Lab Invest1975;33:16-27.
29.
29. Bindal RD, Carlson KE, Katzenellenbogen BS, Katzenellenbogen JA. Lipophilic impurities, not phenolsulfonphthalein, account for the estrogenic activity in commercial preparations of phenol red. J Steroid Biochem1988;31:287-293.
30.
30. Al Saati T, Clamens S, Cohen-Knafo E, et al. Production of monoclonal antibodies to human estrogen-receptor protein (ER) using recombinant ER (RER). Int J Cancer1993;55:651-654.
31.
31. Ryan IP, Schriock ED, Taylor RN, Isolation, characterization and comparison of human endometrial and endometriosis cells in vitro. J Clin endocrinol Metab1994;78:642-649.
32.
32. Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistryh1979;18:5294-5299.
33.
33. Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA. Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci USA1996;93:5925-5930.
34.
34. Mosselman S, Polman J, Dijkema R, ERβ: Identification and characterization of a novel human estrogen receptor. FEBS Lett1996;392:49-53.
35.
35. Sadovsky Y, Riemer RK, Roberts JM. The concentration of estrogen receptors in rabbit uterine myocytes decreases in culture. Am J Obstet Gynecol1992;167:1631-1635.
36.
36. de Ziegler D, Bessis R, Frydman R. Vascular resistance of uterine arteries: Physiological effects of estradiol and progesterone. Fertil Steril1991;55:775-779.
37.
37. Taylor N. Review: Placental sulfatase deficiency. J Inherit Metab Dis1982;5:164-176.
38.
38. Conte FA, Grumbach MM, Ito Y, Fisher CR, Simpson ER. A syndrome of female pseudohermaphrodism, hypergonadotropic hypogonadism, and multicystic ovaries associated with missense mutations in the gene encoding aromatase (P450arom). J Clin Endocrinol Metab1994;78:1287-1292.
39.
39. Smith EP, Boyd J, Frank GR, et al. Estrogen resistance caused by a mutation in the estrogen-receptor gene in a man. N Engl J Med1994;331:1056-1061.
40.
40. Morales DE, McGowan KA, Grant DS, et al. Estrogen promotes angiogenic activity in human umbilical vein endothelial cells in vitro and in a murine model. Circulation1995;91:755-763.
41.
41. Cid MC, Kleinman HK, Grant DS, Schnaper HW, Fauci AS, Hoffman GS. Estradiol enhances leukocyte binding to tumor necrosis factor (TNF)-stimulated endothelial cells via an increase in TNF-induced adhesion molecules E-selectin, intercellular adhesion molecule type 1, and vascular cell adhesion molecule type 1. J Clin Invest1994;93:17-25.
42.
42. Nakai K, Itoh C, Hotta K, Itoh T, Yoshizumi M, Hiramori K. Estradiol-17 beta regulates the induction of VCAM-1 mRNA expression by interleukin-1 beta in human umbilical vein endothelial cells. Life Sci1994;54:PL221-PL227.
43.
43. Morley P, Whitfield JF, Vanderhyden BC, Tsang BR, Schwartz JL. A new, nongenomic estrogen action: The rapid release of intracellular calcium. Endocrinology1992;131:1305-1312.
44.
44. Lieberherr M, Grosse B, Kachkache M, Balsan S. Cell signaling and estrogens in female rat osteoblasts: A possible involvement of unconventional nonnuclear receptors. J Bone Miner Res1993;8:1365-1376.
45.
45. Keresztes P, Ayers JW, Menon KM, Romani T. Comparison of peripheral, uterine and cord estrogen and progesterone levels in laboring and nonlaboring women at term. J Reprod Med1988;33:691-694.
