The mRNA-binding protein HuR is regulated in the menstrual cycle and repressed in ectopic endometrium

Cytokines modulate turnover of the endometrium during the menstrual cycle and contribute to the pathogenesis of endometriosis. Gene expression for cytokines is often regulated by proteins that bind to adenosine- and uridine-rich elements (AREs) in their transcripts to stabilize or destabilize bound messenger RNAs (mRNAs). HuR/ELAVL1 is an RNA-binding protein that stabilizes ARE-containing mRNAs. We hypothesized that HuR might play a role in regulating cytokine expression during the menstrual cycle and in endometriosis and characterized the expression and regulation of HuR in eutopic and ectopic human endometrium. Tissue sections obtained from normal (n = 23) and ectopic (n = 16) endometrium were immunostained for HuR, and staining intensity was evaluated by HSCORE. Cultured stromal cells isolated from normal endometrium were treated with vehicle, estradiol (E2), progesterone (P), E2 + P, tumor necrosis factor-α (TNF-α), and interleukin 1β (IL-1β) for 24 hours, and HuR expression was determined by Western blot. HuR immunoreactivity was significantly lower in the early proliferative and late secretory phases (157.5 ± 11.08 and 190.0 ± 15.2, respectively), compared to the mid-late proliferative (270.0 ± 8.0) and early-mid secretory phases (256.6 ± 20.2; P <.01, analysis of variance [ANOVA]). Furthermore, HuR expression was significantly lower in ectopic endometrial cells compared to normal endometrium in mid-late proliferative and early-mid-secretory phases (P <.01). Estrogen, P, or cytokines did not alter HuR expression in cultured endometrial stromal cells. Increased HuR levels in the mid-menstrual phases are likely to contribute to reduced mid-cycle cytokine expression and enhanced cellular survival in eutopic endometrium. In ectopic endometrium, elevated cytokine levels associated with endometriosis likely reduce HuR expression. Similar content being viewed by others

Hornung D, Ryan IP, Chao VA, Vigne JL, Schriock ED, Taylor RN. Immunolocalization and regulation of the chemokine RANTES in human endometrial and endometriosis tissues and cells. J Clin Endocrinol Metab. 1997;82(5):1621–1628. Kayisli UA, Mahutte N, Arici A. Uterine chemokines in reproductive physiology and pathology. Am J Reprod Immunol. 2002;47(4):213–221. Jones RL, Hannan NJ, Kaitu’u TJ, Zhang J, Salamonsen LA. Identification of chemokines important for leukocyte recruitment to the human endometrim at the times of embryo implnatation and menstruation. J Clin Endocrinol Metab. 2004;89(12):6155–6167. Salamonsen LA, Hannan NJ, Dimitriadis E. Cytokines and chemokines during human embryo implantation: roles in implantation and early placentation. Semin Reprod Med. 2007;25(6):437–444. Wheeler JM. Epidemiology of endometriosis-associated infertility. J Reprod Med. 1989;34(1):41–46. Gao X, Outley J, Botteman M, Spalding J, Simon JA, Pashos CL. Economic burden of endometriosis. Fertil Steril. 2006;86(6):1561–1572. Sampson JA. Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue in peritoneal cavity. Am J Obstet Gynecol. 1927;15:422. Seli E, Arici A. Endometriosis: interaction of immune and endocrine systems. Semin Reprod Med. 2003;21(2):135–144. Seli E, Berkkanoglu M, Arici A. Pathogenesis of endometriosis. Obstet Gynecol Clin N Am. 2003;30(1):41–61. Halme J, Hammond MG, Hulka JF, Raj SG, Talbert LM. Retrograde menstruation in healthy women and in patients with endometriosis. Obstet Gynecol. 1984;64(2):151–154. Liu DT, Hitchcock A. Endometriosis: its association with retrograde menstruation, dysmenorrhea and tubal pathology. Br J Obstet Gynaecol. 1986;93(8):859–862. Fakih H, Bagget B, Holtz G, Tsang KY, Lee JC, Williamson HO. Interleukin-1: possible role in the infertility associated with endometriosis. Fertil Steril. 1987;47(2):213–217. Mori H, Sawairi M, Nakagawa M, Itoh N, Wada K, Tamaya T. Peritoneal fluid interleukin-1 beta and tumor necrosis factor in patients with benign gynecologic disease. Am J Reprod Immunol. 1991;26(2):62–67. Ryan IP, Tseng JF, Schriock ED, Khorram O, Landers DV, Taylor RN. Interleukin-8 concentrations are elevated in peritoneal fluid of women with endometriosis. Fertil Steril. 1995;63(4):929–932. Arici A, Tazuke SI, Attar E, Kliman HJ, Olive DL. Interleukin-8 concentration in peritoneal fluid of patients with endometriosis and modulation of interleukin-8 expression in human mesothelial cells. Mol Hum Reprod. 1996;2(1):40–45. Arici A, Oral E, Attar E, Tazuke S, Olive DL. Monocyte chemotactic protein-1 concentration in peritoneal fluid of women with endometriosis and its modulation of expression in mesothelial cells. Fertil Steril. 1997;67(6):1065–1072. Khorram O, Taylor RN, Ryan IP, Schall TJ, Launders DV. Peritoneal fluid concentrations of the cytokine RANTES correlate with the severity of endometriosis. Am J Obstet Gynecol. 1993;169(6):1545–1549. Eisermann J, Gast MJ, Pineda J, Odem RR, Collins JL. Tumor necrosis factor in peritoneal fluid of women undergoing laparoscopic surgery. Fertil Steril. 1988;50(4):573–579. McLaren J, Prentice A, Charnock-Jones DS, et al. Vascular endothelial growth factor is produced by peritoneal fluid macrophages in endometriosis and is regulated by ovarian steroids. J Clin Invest. 1996;98(2):482–489. Anderson P, Phillips K, Stoecklin G, Kedersha N. Post-transcriptional regulation of pro-infammatory proteins. J Leukoc Biol. 2004;76(1):42–47. Wilusz CJ, Wang W, Peltz SW. Curbing the nonsense: the activation and regulation of mRNA surveillance. Genes Dev. 2001;15(21):2781–2785. Garneau NL, Wilusz J, Wilusz CJ. The highways and byways of mRNA decay. Nat Rev Mol Cell Biol. 2007;8(2):113–126. Espel E. The role of the AU-rich elements of mRNAs in controlling translation. Semin Cell Dev Biol. 2005;16(1):59–67. Wang W, Caldwell MC, Lin S, Furneaux H, Gorospe M. HuR regulates cyclin A and cyclin B1 mRNA stability during cell proliferation. EMBO J. 2000;19(10):2340–2350. Wang W, Furneaux H, Cheng H, et al. HuR regulates p21 mRNA stabilization by UV light. Mol Cell Biol. 2000;20(3):760–769. Chen CY, Xu N, Shyu AB. Highly selective actions of HuR in antagonizing AU-rich element-mediated mRNA destabilization. Mol Cell Biol. 2002;22(20):7268–7278. Atasoy U, Curry SL, Lopez de Silanes I, et al. Regulation of eotaxin gene expression by TNF and IL-4 through messenger RNA stabilization: involvement of the RNA-binding protein HuR. J Immunol. 2003;171(8):4369–4378. Suswam EA, Nabors LB, Huang Y, Yang X, King PH. IL-1beta induces stabilization of IL-8 mRNA in malignant breast cancer cells via the 3’ untranslated region: Involvement of divergent RNA-binding factors HuR, KSRP and TIAR. Int J Cancer. 2005;113(6):911–919. Brennan CM, Steitz JA. HuR and mRNA stability. Cell Mol Life Sci. 2001;58(2):266–277. Palanisamy V, Park NJ, Wang J, Wong DT. AUF1 and HuR proteins stabilize interleukin-8 mRNA in human saliva. J Dent Res. 2008;87(8):772–776. Dixon DA, Tolley ND, King PH, et al. Altered expression of the mRNA stability factor HuR promotes cyclooxygenase-2 expression in colon cancer cells. J Clin Invest. 2001;108(11):1657–1665. Huwiler A, Akool el-S, Aschrafi A, Hamada FM, Pfeilschifter J, Eberhardt W. ATP potentiates interleukin-1 beta-induced MMP-9 expression in mesangial cells via recruitment of the ELAV protein HuR. J Biol Chem. 2003;278(51):51758–51769. Sakai K, Kitagawa Y, Saiki M, Saiki S, Hirose G. Binding of the ELAV-like protein in murine autoimmune T-cells to the nonameric AU-rich element in the 39 untranslated region of CD154 mRNA. Mol Immunol. 2003;39(14):879–883. Blaxall BC, Pellett AC, Wu SC, Pende A, Port JD. Purification and characterization of beta-adrenergic receptor mRNA-binding proteins. J Biol Chem. 2000;275(6):4290–4297. Myer VE, Fan XC, Steitz JA. Identification of HuR as a protein implicated in AUUA-mediated mRNA decay. EMBO J. 1997;16(8):2130–2139. Noyes RW, Hertig AT, Rock J. Dating the endometrial biopsy. Am J Obstet Gynecol. 1975;122(2):262–263. Arici A, Head JR, MacDonald PC, Casey ML. Regulation of interleukin-8 gene expression in human endometrial cells in culture. Mol Cell Endocrinol. 1993;94(2):195–204. American Fertility Society. Revised American Fertility Society classification of endometriosis. Fertil Steril. 1985;43(3):351–352. Ulukus M, Ulukus EC, Tavmergen GEN, Tavmergen E, Zheng W, Arici A. Expression of interleukin-8 and monocyte chemotactic protein 1 in women with endometriosis. Fertil Steril. 2009;91(3):687–693. Seli E, Guzeloglu-Kayisli O, Cakmak H, Kayisli UA, Selam FB, Arici A. Estradiol increases apoptosis in cultured human coronary artery endothelial cells (HCAEC) by up-regulating Fas expression. J Clin Endocrinol Metab. 2006;91(12):4995–5001. Schall TJ, Bacon K, Toy KJ, Goeddel DV. Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES. Nature. 1990;347(6294):669–671. Jones RL, Kelly RW, Critchley HO. Chemokine and cyclooxygenase-2 expression in human endometrium coincides with leukocyte accumulation. Hum Reprod. 1997;12(6):1300–1306. Abdelmohsen K, Lal A, Kim HH, Gorospe M. Post-trascriptional orchestration of an anti-apoptotic program by HuR. Cell Cycle. 2007;6(11):1288–1292. Stewart CL. Leukemia inhibitory factor and the regulation of pre-implantation development of the mammalian embryo. Mol Reprod Dev. 1994;39(2):233–238. Jeyaraj SC, Singh M, Ayupova DA, Govindaraju S, Lee BS. Transcriptional control of human antigen R by bone morphogenic protein. J Biol Chem. 2010;285(7):4432–4440. Kang MJ, Ryu BK, Lee MG, et al. NF-kappaB activates transcription of the RNA-binding factor HuR via PI3K-AKT signaling o promote gastric tumorigenesis. Gastroenterology. 2008;135(6):2030–2042. Farage MA, Neill S, MacLean AB. Physiological changes associated with the menstrual cycle: a review. Obstet Gynecol Surv. 2009;64(1):58–72. Bourdeau V, Deschenes J, Metivier R, et al. Genome-wide identification of high-affinity estrogen response elements in human and mouse. Mol Endocrinol. 2004;18(6):1411–1427. Bourdeau V, Deschenes J, Laperriere D, Aid M, White JH, Mader S. Mechanisms of primary and secondary estrogen target gene regulation in breast cancer cells. Nucleic Acids Res. 2008;36(1):76–93. Frasor J, Danes JM, Komm B, Chang KC, Lyttle CR, Katzenellnbogen BS. Profiling of estrogen up- and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype. Endocrinology. 2003;144(10):4562–4574. Hua S, Kallen CB, Dhar R, et al. Genomic analysis of estrogen cascade reveals histone variant HRA.Z associated with breast cancer progression. Mol Syst Biol. 2008;4:188. Arici A, Seli E, Zeyneloglu HB, Senturk LM, Oral E, Olive DL. Interleukin-8 induces proliferation of endometrial stromal cells: a potential autocrine growth factor. J Clin Endocrinol Metab. 1998;83(4):1201–1205. Akoum A, Lemay A, McColl S, Turcot-Lemay L, Maheux R. Elevated concentration and biologic activity of monocyte chemotactic protein-1 in the peritoneal fluid of patients with endometriosis. Fertil Steril. 1996;66(1):17–23. Khorram O, Taylor RN, Ryan IP, Schall TJ, Landers DV. Peritoneal fluid concentrations of the cytokine RANTES correlate with the severity of endometriosis. Am J Obstet Gynecol. 1993;169(6):1545–1549. Arici A, MacDonald PC, Casey ML. Regulation of monocyte chemotactic protein-1 gene expression in human endometrial cells in cultures. Mol Cell Endocrinol. 1995;107(2):189–197. Lebovic DI, Chao VA, J.F.M, Taylor RN. IL-1 beta induction of RANTES (Regulated upon Activation, Normal T Cell Expressed and Secreted) chemokine gene expression in endometriotic stromal cells depends on a nuclear factor-kB Site in the proximal promoter. J Clin Endocrinol Metab. 2001;86(10):4759–4764. Calaluce R, Gubin MM, Davis JW, et al. The RNA binding protein HuR differentially regulates unique subsets of mRNAs in estrogen receptor negative and estrogen receptor positive breast cancer. BMC Cancer. 2010;10:126. Bhattacharrya SN, Habernacher R, Martine U, Closs EI, Filipoowicz W. Relief of microRNA-mediated translational repression in human cells subjected to stress. Cell. 2006;125(6):1111–1124. Pillai RS, Bhattacharyya SN, Filipowicz W. Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol. 2007;17(3):118–126. Kuokkanen S, Chen B, Ojalvo L, Benard L, Santoro N, Pollard JW. Genomic profiling of microNRAs and messenger RNAs reveals hormonal regulation on microRNA expression in human endometrium. Biol Reprod. 2010;82(4):791–801. Guo X, Wu Y, Hartley RS. MicroRNA-125a represses cell growth by targeting HuR in breast cancer. RNA Biol. 2009;6(5):575–583. Author information Authors and Affiliations Corresponding author Rights and permissions About this article Cite this article Karipcin, F.S., Ensari, T.A., Kayisli, U.A. et al. The mRNA-Binding Protein HuR is Regulated in the Menstrual Cycle and Repressed in Ectopic Endometrium. Reprod. Sci. 18, 145–155 (2011). https://doi.org/10.1177/1933719110382307 Published: Issue date: DOI: https://doi.org/10.1177/1933719110382307