Silencing of SRA1 Regulates ER Expression and Attenuates the Growth of Stromal Cells in Ovarian Endometriosis

Estradiol and its nuclear receptors, estrogen receptor (ER) α and ER-β, have important functions in endometriosis, and the transcriptional activity of these receptors is modulated by coactivators and corepressors. The steroid receptor RNA activator 1 (SRA1) produces SRA long noncoding RNA (lncRNA) and SRA protein (SRAP), which regulate ER expression at the RNA and protein levels in some hormone-dependent tumors via an alternative splicing event. However, only a few are reported on their expressions in endometriosis. Here, we observed that low expression levels of SRA lncRNA and ER-α but relatively high expression levels of SRAP and ER-β were detected in ovarian endometriotic tissues versus normal endometrial tissues. Steroid receptor RNA activator I-small interfering RNA treatment significantly increased ER-α levels but reduced ER-β levels in endometriotic stromal cells (ESCs). Furthermore, the treatment can also attenuate the proliferation and promote early apoptosis in these cells. Our results indicate that the regulation of ER via SRA in ovarian endometriosis may play a significant role in the growth of ESCs. Similar content being viewed by others

Giudice LC, Kao LC. Endometriosis. Lancet. 2004;364(9447):1789–1799. Trukhacheva E, Lin Z, Reierstad S, Cheng Y, Milad M, Bulun SE. Estrogen receptor (ER)-β regulates ER-α expression in stromal cells derived from ovarian endometriosis. J Clin Endocrinol Metab. 2009;94(2):615–622. Lanz RB, McKenna NJ, Onate SA, et al. A steroid receptor coactivator, SRA, functions as an RNA and is present in an SRC-1 complex. Cell. 1999;97(1):17–27. Hube F, Guo J, Chooniedass-Kothari S, et al. Alternative splicing of the first intron of the steroid receptor RNA activator (SRA) participates in the generation of coding and noncoding RNA iso-forms in breast cancer cell lines. DNA Cell Biol. 2006;25(7):418–428. McKay DB, Xi L, Barthel KK, Cech TR. Structure and function of steroid receptor RNA activator protein, the proposed partner of SRA noncoding RNA. J Mol Biol. 2014;426(8):1766–1785. Coleman KM, Lam V, Jaber BM, Lanz RB, Smith CL. SRA coactivation of estrogen receptor-alpha is phosphorylation-independent, and enhances 4-hydroxytamoxifen agonist activity. Biochem Biophys Res Commun. 2004;323(1):332–338. Cooper C, Guo J, Yan Y, et al. Increasing the relative expression of endogenous non-coding Steroid Receptor RNA Activator (SRA) in human breast cancer cells using modified oligonucleo-tides. Nucleic Acids Res. 2009;37(13):4518–4531. Deblois G, Giguere V. Ligand-independent coactivation of ERal-pha AF-1 by steroid receptor RNA activator (SRA) via MAPK activation. J Steroid BiochemMol Biol. 2003;85(2-5):123–131. Klinge CM, Jernigan SC, Mattingly KA, Risinger KE, Zhang J. Estrogen response element-dependent regulation of tran-scriptional activation of estrogen receptors alpha and beta by coactivators and corepressors. J Mol Endocrinol. 2004;33(2):387–410. Watanabe M, Yanagisawa J, Kitagawa H, et al. A subfamily of RNA-binding DEAD-box proteins acts as an estrogen receptor coactivator through the N-terminal activation domain (AF-1) with an RNA coactivator, SRA. EMBOJ. 2001;20(6):1341–1352. Chooniedass-Kothari S, Vincett D, Yan Y, et al. The protein encoded by the functional steroid receptor RNA activator is a new modulator of ER alpha transcriptional activity. FEBS Lett. 2010;584(6):1174–1180. Lin K, Ma J, Wu R, Zhou C, Lin J. Influence of ovarian endome-trioma on expression of steroid receptor RNA activator, estrogen receptors, vascular endothelial growth factor, and thrombospon-din 1 in the surrounding ovarian tissues. Reprod Sci. 2014;21(2):183–189. Lin K, Zhan H, Ma J, et al. Increased steroid receptor RNA activator protein (SRAP) accompanied by decreased estrogen receptor-beta (ER-b) levels during the malignant transformation of endometriosis associated ovarian clear cell carcinoma. Acta Histochem. 2014;116(5):878–882. Jiang QY, Xia JM, Ding HG, Fei XW, Lin J, Wu RJ. RNAi-mediated blocking of ezrin reduces migration of ectopic endome-trial cells in endometriosis. Mol Hum Reprod. 2012;18(9):435–441. Friel AM, Sexton DJ, O’reilly MW, Smith TJ, Morrison JJ. Rho A/Rho kinase: human umbilical artery mRNA expression in normal and preeclamptic pregnancies and functional role in isoprostane-induced vasoconstriction. Reproduction. 2006;132(1):169–176. Zhu R, Xu R, Jiang X, et al. Expression profile of cancer-related genes in human adult bone marrow-derived neural stem like cells highlights the need for tumorigenicity study. J Neurosci Res. 2007;85(14):3064–3070. Pang J, Liu WP, Liu XP, et al. Profiling protein markers associated with lymph node metastasis in prostate cancer by DIGE-based proteomics analysis. J Proteome Res. 2010;9(1):216–226. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C (T)) method. Methods. 2001;25(4):402–408. Chooniedass-Kothari S, Emberley E, Hamedani MK, et al. The steroid receptor RNA activator is the first functional RNA encoding a protein. FEBS Lett. 2004;566(1-3):43–47. Emberley E, Huang GJ, Hamedani MK, et al. Identification of new human coding steroid receptor RNA activator isoforms. Biochem Biophys Res Commun. 2003;301(2):509–515. Li X, Darzynkiewicz Z. Labelling DNA strand breaks with BrdUTP. Detection of apoptosis and cell proliferation. Cell Prolif. 1995;28(11):571–579. Black DL. Mechanisms of alternative pre-messenger RNA splicing. Annu Rev Biochem. 2003;72:291-336. Black DL, Grabowski PJ. Alternative pre-mRNA splicing and neuronal function. Progr Mol Subcell Biol. 2003;31:187-216. Lee CJ, Irizarry K. Alternative splicing in the nervous system: an emerging source of diversity and regulation. Biol Psychiatry. 2003;54(8):771–776. Xue Q, Lin Z, Cheng YH, et al. Promoter methylation regulates estrogen receptor 2 in human endometrium and endometriosis. Biol Reprod. 2007;77(4):681–687. Fazleabas AT, Brudney A, Chai D, Langoi D, Bulun SE. Steroid receptor and aromatase expression in baboon endometriotic lesions. Fertil Steril. 2003;80(suppl 2):820–827. Hube F, Velasco G, Rollin J, Furling D, Francastel C. Steroid receptor RNA activator protein binds to and counteracts SRA RNA-mediated activation of MyoD and muscle differentiation. Nucleic Acids Res. 2011;39(2):513–525. Cooper C, Vincett D, Yan Y, Hamedani MK, Myal Y, Leygue E. Steroid receptor RNA activator bi-faceted genetic system: heads or tails? Biochimie. 2011;93(11):1973–1980. Lanz RB, Razani B, Goldberg AD, O’Malley BW. Distinct RNA motifs are important for coactivation of steroid hormone receptors by steroid receptor RNA activator (SRA). Proc Natl Acad Sci USA. 2002;99(25):16081–16086. Zhao X, Patton JR, Davis SL, Florence B, Ames SJ, Spanjaard RA. Regulation of nuclear receptor activity by a pseudouridine synthase through posttranscriptional modification of steroid receptor RNA activator. Mol Cell. 2004;15(4):549–558. Borth N, Massier J, Franke C, Sachse K, Saluz HP, Hanel F. Chlamydial protease CT441 interacts with SRAP1 co-activator of estrogen receptor alpha and partially alleviates its co-activation activity. J Steroid Biochem Mol Biol. 2010;119(1-2): 89–95. Kawashima H, Takano H, Sugita S, Takahara Y, Sugimura K, Nakatani T. A novel steroid receptor co-activator protein (SRAP) as an alternative form of steroid receptor RNA-activator gene: expression in prostate cancer cells and enhancement of androgen receptor activity. Biochem J. 2003;369(pt 1):163–171. Kurisu T, Tanaka T, Ishii J, et al. Expression and function of human steroid receptor RNA activator in prostate cancer cells: role of endogenous hSRA protein in androgen receptor-mediated transcription. Prostate Cancer Prostatic Dis. 2006;9(2):173–178. Chooniedass-Kothari S, Hamedani MK, Auge C, et al. The steroid receptor RNA activator protein is recruited to promoter regions and acts as a transcriptional repressor. FEBS Lett. 2010;584(11):2218–2224. Bulun SE. Endometriosis. N Engl J Med. 2009;360(3):268–279. Di Ruscio A, Ebralidze AK, Benoukraf T, et al. DNMT1-interacting RNAs block gene-specific DNA methylation. Nature. 2013;503(7476):371–376. Arab K, Park YJ, Lindroth AM, et al. Long noncoding RNA TARID directs demethylation and activation of the tumor suppressor TCF21 via GADD45A. Mol Cell. 2014;55(4):604–614. Leygue E. Steroid receptor RNA activator (SRA1): unusual bifa-ceted gene products with suspected relevance to breast cancer. Nucl Recept Signal. 2007;5:e006. Foulds CE, Tsimelzon A, Long W, et al. Research resource: expression profiling reveals unexpected targets and functions of the human steroid receptor RNA activator (SRA) gene. Mol Endocrinol. 2010;24(5):1090–1105. Yan Y, Skliris GP, Penner C, et al. Steroid Receptor RNA Activator Protein (SRAP): a potential new prognostic marker for estrogen receptor-positive/node-negative/younger breast cancer patients. Breast Cancer Res. 2009;11(5):R67. Han SJ, Jung SY, Wu SP, et al. Estrogen receptor beta modulates apoptosis complexes and the inflammasome to drive the patho-genesis of endometriosis. Cell. 2015;163(4):960–974. Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Lin, K., Zhan, H., Ma, J. et al. Silencing of SRA1 Regulates ER Expression and Attenuates the Growth of Stromal Cells in Ovarian Endometriosis. Reprod. Sci. 24, 836–843 (2017). https://doi.org/10.1177/1933719116670036 Published: Issue date: DOI: https://doi.org/10.1177/1933719116670036