Effect of In vitro Culture of Human Myometrial and Fibroid Smooth Muscle Cells on Protein Estrogen Receptor

Main Article Content

Duangporn Saengwimol
Wanwisa Waiyaput
Somphoch Pumipichet
Areepan Sophonsritsuk

Abstract

Background: Uterine leiomyomas (fibroids, myomas) are the most common benign smooth muscle cell tumors in the reproductive tract. The sex hormone estrogen has been implicated as a major regulator of fibroid growth. Estrogen exerts its physiological effects on the target cells by binding to specific nuclear receptors, the estrogen receptor α (ERα) and estrogen receptor ß (ERß).


Objective: To assess the expression of ERα and progesterone receptor (PR) on primary culture smooth muscle cells (SMCs) from myometrial and fibroid tissue for the potential use of primary culture SMCs as an experimental model for studying ER activity, PR activity, and drug testing.


Methods: Quantitative real-time PCR (qPCR) and Western blot analysis were used to compare ERα  between paired myometrial and fibroid tissues, as well as cells isolated from the same tissues and cultured up to the three passages.


Results: ERα and PR levels in cultured cells were rapidly lost comparing with whole tissues for both myometrium and fibroid. Moreover, ERα protein was degraded after tissue digestion prior to the culturing process.


Conclusions: Primary human myometrial and fibroid smooth muscle cell cultures are not suitable for studying ER in vitro model. The stable ER and PR cell lines were the model of choice. 


 

Article Details

How to Cite
1.
Saengwimol D, Waiyaput W, Pumipichet S, Sophonsritsuk A. Effect of In vitro Culture of Human Myometrial and Fibroid Smooth Muscle Cells on Protein Estrogen Receptor. Rama Med J [Internet]. 2016 Dec. 30 [cited 2024 Mar. 29];39(4):206-1. Available from: https://he02.tci-thaijo.org/index.php/ramajournal/article/view/80053
Section
Original Articles

References

Verkauf BS. Myomectomy for fertility enhancement and preservation. Fertil Steril. 1992;58:1-15.

Baird DD, Dunson DB, Hill MC, Cousins D, Schectman JM. High cumulative incidence of uterine leiomyoma in black and white women: ultrasound evidence. Am J Obstet Gynecol. 2003;188:100-7.

Bakas P, Liapis A, Vlahopoulos S, Giner M, Logotheti S, Creatsas G, et al. Estrogen receptor alpha and beta in uterine fibroids: a basis for altered estrogen responsiveness. Fertil Steril. 2008;90:1878-85.

Flake GP, Andersen J, Dixon D. Etiology and pathogenesis of uterine leiomyomas: a review. Environ Health Perspect. 2003;111:1037-54.

Swartz CD, Afshari CA, Yu L, Hall KE, Dixon D. Estrogen-induced changes in IGF-I, Myb family and MAP kinase pathway genes in human uterine leiomyoma and normal uterine smooth muscle cell lines. Mol Hum Reprod. 2005;11:441-50.

Sadan O, van Iddekinge B, van Gelderen CJ, Savage N, Becker PJ, van der Walt LA, et al. Oestrogen and progesterone receptor concentrations in leiomyoma and normal myometrium. Ann Clin Biochem. 1987;24:263-7.

Tamaya T, Fujimoto J, Okada H. Comparison of cellular levels of steroid receptors in uterine leiomyoma and myometrium. Acta Obstet Gynecol Scand. 1985;64:307-9.

Raz L, Khan MM, Mahesh VB, Vadlamudi RK, Brann DW. Rapid estrogen signaling in the brain. Neurosignals. 2008;16:140-53.

Rissman EF, Wersinger SR, Taylor JA, Lubahn DB. Estrogen receptor function as revealed by knockout studies: neuroendocrine and behavioral aspects. Horm Behav. 1997;31:232-43.

Gargett CE, Zaitseva M, Bucak K, Chu S, Fuller PJ, Rogers PA. 17Beta-estradiol up-regulates vascular endothelial growth factor receptor-2 expression in human myometrial microvascular endothelial cells: role of estrogen receptor-alpha and -beta. J Clin Endocrinol Metab. 2002;87:4341-9.

Gargett CE, Bucak K, Zaitseva M, Chu S, Taylor N, Fuller PJ, et al. Estrogen receptor-alpha and -beta expression in microvascular endothelial cells and smooth muscle cells of myometrium and leiomyoma. Mol Hum Reprod. 2002;8:770-5.

Sadovsky Y, Riemer RK, Roberts JM. The concentration of estrogen receptors in rabbit uterine myocytes decreases in culture. Am J Obstet Gynecol. 1992;167:1631-5.

Severino MF, Murray MJ, Brandon DD, Clinton GM, Burry KA, Novy MJ. Rapid loss of oestrogen and progesterone receptors in human leiomyoma and myometrial explant cultures. Mol Hum Reprod. 1996;2:823-8.

Zaitseva M, Vollenhoven BJ, Rogers PA. In vitro culture significantly alters gene expression profiles and reduces differences between myometrial and fibroid smooth muscle cells. Mol Hum Reprod. 2006;12:187-207.

Alarid ET, Bakopoulos N, Solodin N. Proteasome-mediated proteolysis of estrogen receptor: a novel component in autologous down-regulation. Mol Endocrinol. 1999;13:1522-34.

Alarid ET. Lives and times of nuclear receptors. Mol Endocrinol. 2006;20:1972-81.

Valley CC, Solodin NM, Powers GL, Ellison SJ, Alarid ET. Temporal variation in estrogen receptor-alpha protein turnover in the presence of estrogen. J Mol Endocrinol. 2008;40:23-34.

Blake RE. Leiomyomata uteri: hormonal and molecular determinants of growth. J Natl Med Assoc. 2007;99:1170-84.

Maruo T, Ohara N, Yoshida S, Nakabayashi K, Sasaki H, Xu Q, et al. Translational research with progesterone receptor modulator motivated by the use of levonorgestrel-releasing intrauterine system. Contraception. 2010;82:435-41.

Soules MR, McCarty KS, Jr. Leiomyomas: steroid receptor content. Variation within normal menstrual cycles. Am J Obstet Gynecol. 1982;143:6-11.

Wilson EA, Yang F, Rees ED. Estradiol and progesterone binding in uterine leiomyomata and in normal uterine tissues. Obstet Gynecol. 1980;55:20-4.

Sampath D, Zhu Y, Winneker RC, Zhang Z. Aberrant expression of Cyr61, a member of the CCN (CTGF/Cyr61/Cef10/NOVH) family, and dysregulation by 17 beta-estradiol and basic fibroblast growth factor in human uterine leiomyomas. J Clin Endocrinol Metab 2001;86:1707-15.

Wijayaratne AL, McDonnell DP. The human estrogen receptor-alpha is a ubiquitinated protein whose stability is affected differentially by agonists, antagonists, and selective estrogen receptor modulators. J Biol Chem. 2001;276:35684-92.

Harris DM, Besselink E, Henning SM, Go VL, Heber D. Phytoestrogens induce differential estrogen receptor alpha- or Beta-mediated responses in transfected breast cancer cells. Exp Biol Med (Maywood). 2005;230:558-68.

Mangioni S, Vigano P, Lattuada D, Abbiati A, Vignali M, Di Blasio AM. Overexpression of the Wnt5b gene in leiomyoma cells: implications for a role of the Wnt signaling pathway in the uterine benign tumor. J Clin Endocrinol Metab. 2005;90:5349-55.

Carney SA, Tahara H, Swartz CD, Risinger JI, He H, Moore AB, et al. Immortalization of human uterine leiomyoma and myometrial cell lines after induction of telomerase activity: molecular and phenotypic characteristics. Lab Invest. 2002;82:719-28.

Varella-Garcia M, Chen L, Zheng X, Yu L, Dixon D. Karyotypic characteristics of human uterine leiomyoma and myometrial cell lines following telomerase induction. Cancer Genet Cytogenet. 2006;170:71-5.