Coronary artery and cultured aortic smooth muscle cells express mRNA for both the classical estrogen receptor and the newly described estrogen receptor beta

Estrogens exhibit potent anti-atherogenic effects through mechanisms which may involve direct effects on the artery. The existence of the classical estrogen receptor (ERalpha) in vascular tissues has been established. Recently a new estrogen receptor (ERbeta) has been discovered which represents a distinct gene product with homology to the classical ERalpha. The purpose of the present study was to determine if ERbeta mRNA is expressed in vascular tissues of female and male primates. Oligonucleotide primers were developed for the specific RT-PCR amplification of ERalpha or ERbeta mRNA. RT-PCR products of the appropriate size for ERalpha and for ERbeta were observed after amplification of RNA isolated from coronary arteries of both male and female cynomolgus monkeys. Similar results were obtained from cultured aortic smooth muscle cells and from monkey reproductive tissues such as ovary and uterus. The relative expression of ERbeta to ERalpha mRNA was greatest in ovary, on the same order of magnitude in monkey vascular tissues and uterus, while the human breast cancer cell line MCF-7 exhibited a very low level of ERbeta relative to ERalpha. Sequence analysis of isolated RT-PCR products showed >95% similarity between the monkey and the published human sequences for both ERalpha and ERbeta. These findings suggest that estrogen may influence vascular gene expression not only through classical ERalpha but also through the newly described ERbeta. These findings also demonstrate the potential for targeting of these receptors in males for prevention or treatment of heart disease.

[1]  H. Mcgill,et al.  Hormone Receptors of the Baboon Cardiovascular System: Biochemical Characterization of Aortic and Myocardial Cytoplasmic Progesterone Receptors , 1982, Circulation research.

[2]  Michael V. Doyle,et al.  Quantitation of mRNA by the Polymerase Chain Reaction , 1989 .

[3]  J. Polman,et al.  ERβ: Identification and characterization of a novel human estrogen receptor , 1996 .

[4]  Y. Ouchi,et al.  Vascular smooth muscle cells as target for estrogen. , 1993, Biochemical and biophysical research communications.

[5]  J. Gustafsson,et al.  Cloning of a novel receptor expressed in rat prostate and ovary. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[6]  S. Murota,et al.  Estradiol-binding sites in rat aortic smooth muscle cells in culture. , 1981, Atherosclerosis.

[7]  K. Grandien,et al.  Printed in U.S.A. Copyright © 1997 by The Endocrine Society Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors � and � , 2022 .

[8]  Estrogen and coronary heart disease in women. , 1991 .

[9]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[10]  C. Shively,et al.  From menarche to menopause: coronary artery atherosclerosis and protection in cynomolgus monkeys. , 1989, American journal of obstetrics and gynecology.

[11]  J. Shine,et al.  Sequence and expression of human estrogen receptor complementary DNA. , 1986, Science.

[12]  R. Karas,et al.  Human vascular smooth muscle cells express an estrogen receptor isoform , 1995, FEBS letters.

[13]  T. Clarkson,et al.  Effects of oestrogens and progestogens on coronary atherosclerosis and osteoporosis of monkeys. , 1991, Bailliere's clinical obstetrics and gynaecology.

[14]  D. Harder,et al.  Estrogen receptors and effects of estrogen on membrane electrical properties of coronary vascular smooth muscle , 1979, Journal of cellular physiology.

[15]  P. Chomczyński,et al.  A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. , 1993, BioTechniques.

[16]  J. Gustafsson,et al.  Estrogen receptor-beta mRNA expression in rat ovary: down-regulation by gonadotropins. , 1997, Molecular endocrinology.

[17]  J. Isner,et al.  Variable expression of the estrogen receptor in normal and atherosclerotic coronary arteries of premenopausal women. , 1994, Circulation.

[18]  H. Mcgill,et al.  Nuclear Uptake of Sex Steroid Hormones in the Cardiovascular System of the Baboon , 1981, Circulation research.

[19]  S. Manuck,et al.  Inhibition of coronary artery atherosclerosis by 17-beta estradiol in ovariectomized monkeys. Lack of an effect of added progesterone. , 1990, Arteriosclerosis.

[20]  G. Delsol,et al.  Estrogen synthesis, estrogen metabolism, and functional estrogen receptors in rat arterial smooth muscle cells in culture. , 1995, Endocrinology.

[21]  H. Mcgill,et al.  Hormone Receptors of the Baboon Cardiovascular System , 1981, Arteriosclerosis.

[22]  B. Katzenellenbogen,et al.  Proliferation, hormonal responsiveness, and estrogen receptor content of MCF-7 human breast cancer cells grown in the short-term and long-term absence of estrogens. , 1987, Cancer research.

[23]  H. S. Klopfenstein,et al.  Estrogen modulates responses of atherosclerotic coronary arteries. , 1990, Circulation.

[24]  K. Horwitz,et al.  Canine vascular tissues are targets for androgens, estrogens, progestins, and glucocorticoids. , 1982, The Journal of clinical investigation.

[25]  S. Shain,et al.  Estradiol‐I 78 Affects Estrogen Receptor Distribution and Elevates Progesterone Receptor Content in Baboon Aorta , 1986, Arteriosclerosis.

[26]  P. Colburn,et al.  Estrogen-binding sites in endothelial cell cultures , 1978 .

[27]  A. Orekhov,et al.  Smooth Muscle Cells from Adult Human Aorta , 1990 .

[28]  M. Adams,et al.  Medroxyprogesterone acetate antagonizes inhibitory effects of conjugated equine estrogens on coronary artery atherosclerosis. , 1997, Arteriosclerosis, thrombosis, and vascular biology.

[29]  R. Karas,et al.  Human vascular smooth muscle cells contain functional estrogen receptor. , 1994, Circulation.