17β-Estradiol Induces Overproliferation in Adenomyotic Human Uterine Smooth Muscle Cells of the Junctional Zone Through Hyperactivation of the Estrogen Receptor-Enhanced RhoA/ROCK Signaling Pathway

Adenomyosis (ADS) is a common estrogen-dependent gynecological disease with unknown etiology. Recent models favor abnormal thickening of the junctional zone (JZ) may be the causative factor in the development of ADS. RhoA, a small guanosine triphosphatase which controls multiple cellular processes, is involved in the control of cell proliferation. Here we demonstrate that treatment of human uterine smooth muscle cells (SMCs) of the JZ with 17β-estradiol (E2) increased expression of RhoA and its downstream effectors (-associated coiled coil containing protein kinase [ROCK] 1 and ROCK2). Compared with non-ADS cells, RhoA, ROCK1, and ROCK2 were overexpressed and hyperactivated in ADS cells. These effects were suppressed in the presence of ICI 182,780, supporting an estrogen receptor (ER)-dependent mechanism. Hyperactivation of ER-enhanced RhoA/ROCK signaling was associated with overproliferation in ADS human uterine SMCs of the JZ. Moreover, E2-induced overproliferation was accompanied by downregulation of cyclin-dependent kinases inhibitors (CKIs; p21Waf1/Cip1 and p27Kip1) and upregulation of cyclin-dependent kinases (CDKs) and cyclins (cyclin D1, cyclin E1, CDK2, CDK4, and CDK6). Similar content being viewed by others

Bird CC, McElin TW, Manalo-Estrella P. The elusive adenomyosis of the uterus—revisited. Am J Obstet Gynecol. 1972;112(5): 583–593. Taran FA, Stewart EA, Brucker S. Adenomyosis: epidemiology, risk factors, clinical phenotype and surgical and interventional alternatives to hysterectomy. Geburtshilfe Frauenheilkd. 2013; 73(9):924–931. Brosens I, Kunz G, Benagiano G. Is adenomyosis the neglected phenotype of an endomyometrial dysfunction syndrome? Gynecol Surg. 2012;9(2):131–137. Novellas S, Chassang M, Delotte J, et al. MRI characteristics of the uterine junctional zone: from normal to the diagnosis of adenomyosis. AJR Am J Roentgenol. 2011;196(5):1206–1213. Buchsbaum RJ. Rho activation at a glance. J Cell Sci. 2007; 120(7):1149–1152. Bryan BA, Dennstedt E, Mitchell DC, et al. RhoA/ROCK signaling is essential for multiple aspects of VEGF-mediated angiogenesis. FASEB J. 2010;24(9):3186–3195. Olson MF. Applications for ROCK kinase inhibition. Curr Opin Cell Biol. 2008;20(2):242–248. Wang S, Duan H, Zhang Y, Liu JJ. Expression of RhoA and Rho kinase in junctional zone of human adenomyosis and its relationship with dysmenorrheal. Zhonghua Fu Chan Ke Za Zhi. 2013; 48(12):911–915. Oviedo PJ, Sobrino A, Laguna-Fernandez A, et al. Estradiol induces endothelial cell migration and proliferation through estrogen receptor-enhanced RhoA/ROCK pathway. Mol Cell Endocrinol. 2011;335(2):96–103. Whirledge S, Dixon D, Cidlowski JA. Glucocorticoids regulate gene expression and repress cellular proliferation in human uterine leiomyoma cells. Horm Cancer. 2012;3(3):79–92. Levy G, Dehaene A, Laurent N, et al. An update on adenomyosis. Diagn Intervent Imaging. 2013;94(1):3–25. Schweppe KW. The place of dydrogesterone in the treatment of endometriosis and adenomyosis. Maturitas. 2009;65(1):S23–S27. Cho S, Nam A, Kim H, et al. Clinical effects of the levonorgestrel-releasing intrauterine device in patients with adenomyosis. Am J Obstet Gynecol. 2008;198(4):371–373. Wang J, Zhang HH, Duan H. Expression of ERalpha in endometrial–myometria interface of human adenomyosis. Zhonghua Yi Xue Za Zhi. 2010;90(27):1914–1917. Zhang S, Tang Q, Xu F, et al. RhoA regulates G1-S progression of gastric cancer cells by modulation of multiple INK4 family tumor suppressors. Mol Cancer Res. 2009;7(4):570–580. Liu X, Chen D, Liu G. Overexpression of RhoA promotes the proliferation and migration of cervical cancer cells. Biosci Biotechnol Biochem. 2014;78(11):1895–1901. Abe H, Kamai T, Hayashi K, et al. The Rho-kinase inhibitor HA-1077 suppresses proliferation/migration and induces apoptosis of urothelial cancer cells. BMC Cancer. 2014;14:412. Tsuno A, Nasu K, Kawano Y, et al. Fasudil inhibits the proliferation and contractility and induces cell cycle arrest and apoptosis of human endometriotic stromal cells: a promising agent for the treatment of endometriosis. J Clin Endocrinol Metab. 2011;96(12):E1944–E1952. Zohrabian VM, Forzani B, Chau Z, et al. Rho/ROCK and MAPK signaling pathways are involved in glioblastoma cell migration and proliferation. Anticancer Res. 2009;29(1): 119–123. Sinnett-Smith J, Lunn JA, Leopoldt D, Rozengurt E. Y-27632, an inhibitor of Rho-associated kinases, prevents tyrosine phosphorylation of focal adhesion kinase and paxillin induced by bombesin: dissociation from tyrosine phosphorylation of p130(CAS). Exp Cell Res. 2001;266(2):292–302. Gerard C, Goldbeter A. The balance between cell cycle arrest and cell proliferation: control by the extracellular matrix and by contact inhibition. Interface Focus. 2014;4(3):20130075. Jiang W, Wang Q, Chen S, et al. Influenza A virus NS1 induces G0/G1 cell cycle arrest by inhibiting the expression and activity of RhoA protein. J Virol. 2013;87(6):3039–3052. Lee MH, Cho YS, Han YM. Simvastatin suppresses self-renewal of mouse embryonic stem cells by inhibiting RhoA geranylgeranylation. Stem Cells. 2007;25(7):1654–1663. Yu L, Quinn DA, Garg HG, Hales CA. Heparin inhibits pulmonary artery smooth muscle cell proliferation through guanine nucleotide exchange factor-H1/RhoA/Rho kinase/p27. Am J Respir Cell Mol Biol. 2011;44(4):524–530. Olson MF, Paterson HF, Marshall CJ. Signals from Ras and Rho GTPases interact to regulate expression of p21Waf1/Cip1. Nature. 1998;394(6690):295–299. Zuckerbraun BS, Shapiro RA, Billiar TR, Tzeng E. RhoA influences the nuclear localization of extracellular signal-regulated kinases to modulate p21Waf/Cip1 expression. Circulation. 2003;108(7):876–881. Morgan DO. Principles of CDK regulation. Nature. 1995; 374(6518):131–134. Liberto M, Cobrinik D, Minden A. Rho regulates p21 (CIP1), cyclin D1, and checkpoint control in mammary epithelial cells. Oncogene. 2002;21(10):1590–1599. Viallard JF, Lacombe F, Belloc F, et al. Molecular mechanisms controlling the cell cycle: fundamental aspects and implications for oncology. Cancer Radiother. 2001;5(2): 109–129. Mehasseb MK, Panchal R, Taylor AH, et al. Estrogen and progesterone receptor isoform distribution through the menstrual cycle in uteri with and without adenomyosis. Fertil Steril. 2011;95(7):2228–2235.e1. Nie J, Lu Y, Liu X, Guo SW. Immunoreactivity of progesterone receptor isoform B, nuclear factor kappaB, and IkappaBalpha in adenomyosis. Fertil Steril. 2009;92(3):886–889. Jichan N, Xishi L, Guo SW. Promoter hypermethylation of progesterone receptor isoform B (PR-B) in adenomyosis and its rectification by a histone deacetylase inhibitor and a demethylation agent. Reprod Sci. 2010;17(11):995–1005. Wood C. Surgical and medical treatment of adenomyosis. Hum Reprod Update. 1998;4(4):323–336. Garcia L, Isaacson K. Adenomyosis: review of the literature. J Minim Invasive Gynecol. 2011;18(4):428–437. Farquhar C, Brosens I. Medical and surgical management of adenomyosis. Best Pract Res Clin Obstet Gynaecol. 2006;20(4):603–616. Yamanaka A, Kimura F, Kishi Y, et al. Progesterone and synthetic progestin, dienogest, induce apoptosis of human primary cultures of adenomyotic stromal cells. Eur J Obstet Gynecol Reprod Biol. 2014;179:170–174. Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Sun, FQ., Duan, H., Wang, S. et al. 17β-Estradiol Induces Overproliferation in Adenomyotic Human Uterine Smooth Muscle Cells of the Junctional Zone Through Hyperactivation of the Estrogen Receptor-Enhanced RhoA/ROCK Signaling Pathway. Reprod. Sci. 22, 1436–1444 (2015). https://doi.org/10.1177/1933719115584447 Published: Issue date: DOI: https://doi.org/10.1177/1933719115584447