Advertisement

Journal of Assisted Reproduction and Genetics

, Volume 36, Issue 2, pp 211–221 | Cite as

MiRNA-99a can regulate proliferation and apoptosis of human granulosa cells via targeting IGF-1R in polycystic ovary syndrome

  • Yudi Geng
  • Cong Sui
  • Yang Xun
  • Qiaohong LaiEmail author
  • Lei JinEmail author
Reproductive Physiology and Disease
  • 124 Downloads

Abstract

Purpose

We aimed to evaluate the regulation of miR-99a to the biological functions of granulosa cells in polycystic ovary syndrome (PCOS) via targeting IGF-1R.

Methods

We collected aspirated follicular fluid in both patients with and without PCOS. Granulosa cells (GCs) were isolated through Percoll differential centrifugation to detect both miR-99a and IGF-1R expressions. We further transfected COV434 cells with miR-99a mimics to establish a miRNA-99a (miR-99a) overexpression model. We explored the regulation of miR-99a to the proliferation and apoptosis of human GCs via IGF-1R in COV434. The effect of different insulin concentrations on miR-99a expression was also evaluated.

Results

MiR-99a was significantly downregulated while IGF-1R was upregulated in patients with PCOS. MiR-99a can regulate IGF-1R on a post-transcriptional level. After transfection of miR-99a mimics, the proliferation rate was decreased and apoptosis rate was increased significantly in COV434. Exogenous insulin-like growth factor 1 (IGF-1) treatment could reverse the effect of miR-99a. MiR-99a was negatively and dose-dependently regulated by insulin in vitro.

Conclusions

MiR-99a expression was downregulated in patients with PCOS, the degree of which may be closely related to insulin resistance and hyperinsulinemia. MiR-99a could attenuate proliferation and promote apoptosis of human GCs through targeting IGF-1R, which could partly explain the abnormal folliculogenesis in PCOS.

Keywords

miR-99a Granulosa cell IGF-1R Proliferation Apoptosis Polycystic ovary syndrome 

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 81401268).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    Dalmay T. MicroRNAs and cancer. J Intern Med. 2008;263(4):366–75.  https://doi.org/10.1111/j.1365-2796.2008.01926.x.CrossRefPubMedGoogle Scholar
  2. 2.
    Bartel DP. MicroRNAs: target recognition and regulatory functions. Cell. 2009;136(2):215–33.  https://doi.org/10.1016/j.cell.2009.01.002.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    McKenna LB, Schug J, Vourekas A, McKenna JB, Bramswig NC, Friedman JR, et al. MicroRNAs control intestinal epithelial differentiation, architecture, and barrier function. Gastroenterology. 2010;139(5):1654–64, 64 e1.  https://doi.org/10.1053/j.gastro.2010.07.040.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Fan C, Lin Y, Mao Y, Huang Z, Liu AY, Ma H, et al. MicroRNA-543 suppresses colorectal cancer growth and metastasis by targeting KRAS, MTA1 and HMGA2. Oncotarget. 2016;7(16):21825–39.  https://doi.org/10.18632/oncotarget.7989.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Ge X, Liu X, Lin F, Li P, Liu K, Geng R, et al. MicroRNA-421 regulated by HIF-1alpha promotes metastasis, inhibits apoptosis, and induces cisplatin resistance by targeting E-cadherin and caspase-3 in gastric cancer. Oncotarget. 2016;7(17):24466–82.  https://doi.org/10.18632/oncotarget.8228.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Shukla GC, Singh J, Barik S. MicroRNAs: processing, maturation, target recognition and regulatory functions. Mol Cell Pharmacol. 2011;3(3):83–92.PubMedPubMedCentralGoogle Scholar
  7. 7.
    Mei LL, Qiu YT, Huang MB, Wang WJ, Bai J, Shi ZZ. MiR-99a suppresses proliferation, migration and invasion of esophageal squamous cell carcinoma cells through inhibiting the IGF1R signaling pathway. Cancer Biomark. 2017;20:527–37.  https://doi.org/10.3233/CBM-170345.CrossRefPubMedGoogle Scholar
  8. 8.
    Wang X, Li Y, Qi W, Zhang N, Sun M, Huo Q, et al. MicroRNA-99a inhibits tumor aggressive phenotypes through regulating HOXA1 in breast cancer cells. Oncotarget. 2015;6(32):32737–47.  https://doi.org/10.18632/oncotarget.5355.PubMedPubMedCentralGoogle Scholar
  9. 9.
    Xing B, Ren C. Tumor-suppressive miR-99a inhibits cell proliferation via targeting of TNFAIP8 in osteosarcoma cells. Am J Transl Res. 2016;8(2):1082–90.PubMedPubMedCentralGoogle Scholar
  10. 10.
    Maugeri M, Barbagallo D, Barbagallo C, Banelli B, Di Mauro S, Purrello F, et al. Altered expression of miRNAs and methylation of their promoters are correlated in neuroblastoma. Oncotarget. 2016;7(50):83330–41.  https://doi.org/10.18632/oncotarget.13090.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Weroha SJ, Haluska P. The insulin-like growth factor system in cancer. Endocrinol Metab Clin N Am. 2012;41(2):335–50, vi.  https://doi.org/10.1016/j.ecl.2012.04.014.CrossRefGoogle Scholar
  12. 12.
    Goodarzi MO, Dumesic DA, Chazenbalk G, Azziz R. Polycystic ovary syndrome: etiology, pathogenesis and diagnosis. Nat Rev Endocrinol. 2011;7(4):219–31.  https://doi.org/10.1038/nrendo.2010.217.CrossRefPubMedGoogle Scholar
  13. 13.
    Moran L, Teede H. Metabolic features of the reproductive phenotypes of polycystic ovary syndrome. Hum Reprod Update. 2009;15(4):477–88.  https://doi.org/10.1093/humupd/dmp008.CrossRefPubMedGoogle Scholar
  14. 14.
    Eek D, Paty J, Black P, Celeste Elash CA, Reaney M. A comprehensive disease model of polycystic ovary syndrome (Pcos). Value Health. 2015;18(7):A722.  https://doi.org/10.1016/j.jval.2015.09.2739.CrossRefPubMedGoogle Scholar
  15. 15.
    Diamanti-Kandarakis E, Piperi C, Spina J, Argyrakopoulou G, Papanastasiou L, Bergiele A, et al. Polycystic ovary syndrome: the influence of environmental and genetic factors. Hormones. 2006;5(1):17–34.CrossRefPubMedGoogle Scholar
  16. 16.
    Artimani T, Saidijam M, Aflatoonian R, Amiri I, Ashrafi M, Shabab N, et al. Estrogen and progesterone receptor subtype expression in granulosa cells from women with polycystic ovary syndrome. Gynecol Endocrinol. 2015;31(5):379–83.  https://doi.org/10.3109/09513590.2014.1001733.CrossRefPubMedGoogle Scholar
  17. 17.
    Stubbs SA, Stark J, Dilworth SM, Franks S, Hardy K. Abnormal preantral folliculogenesis in polycystic ovaries is associated with increased granulosa cell division. J Clin Endocrinol Metab. 2007;92(11):4418–26.  https://doi.org/10.1210/jc.2007-0729.CrossRefPubMedGoogle Scholar
  18. 18.
    Das M, Djahanbakhch O, Hacihanefioglu B, Saridogan E, Ikram M, Ghali L, et al. Granulosa cell survival and proliferation are altered in polycystic ovary syndrome. J Clin Endocrinol Metab. 2008;93(3):881–7.  https://doi.org/10.1210/jc.2007-1650.CrossRefPubMedGoogle Scholar
  19. 19.
    Chang RJ, Cook-Andersen H. Disordered follicle development. Mol Cell Endocrinol. 2013;373(1–2):51–60.  https://doi.org/10.1016/j.mce.2012.07.011.CrossRefPubMedGoogle Scholar
  20. 20.
    Yu YS, Sui HS, Han ZB, Li W, Luo MJ, Tan JH. Apoptosis in granulosa cells during follicular atresia: relationship with steroids and insulin-like growth factors. Cell Res. 2004;14(4):341–6.  https://doi.org/10.1038/sj.cr.7290234.CrossRefPubMedGoogle Scholar
  21. 21.
    Cakir E, Topaloglu O, Colak Bozkurt N, Karbek Bayraktar B, Gungunes A, Sayki Arslan M, et al. Insulin-like growth factor 1, liver enzymes, and insulin resistance in patients with PCOS and hirsutism. Turkish J Med Sci. 2014;44(5):781–6.CrossRefGoogle Scholar
  22. 22.
    Hossain MM, Cao M, Wang Q, Kim JY, Schellander K, Tesfaye D, et al. Altered expression of miRNAs in a dihydrotestosterone-induced rat PCOS model. J Ovarian Res. 2013;6(1):36.  https://doi.org/10.1186/1757-2215-6-36.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Roth LW, McCallie B, Alvero R, Schoolcraft WB, Minjarez D, Katz-Jaffe MG. Altered microRNA and gene expression in the follicular fluid of women with polycystic ovary syndrome. J Assist Reprod Genet. 2014;31(3):355–62.  https://doi.org/10.1007/s10815-013-0161-4.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Liu S, Zhang X, Shi C, Lin J, Chen G, Wu B, et al. Altered microRNAs expression profiling in cumulus cells from patients with polycystic ovary syndrome. J Transl Med. 2015;13:238.  https://doi.org/10.1186/s12967-015-0605-y.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Zhang ZW, Guo RW, Lv JL, Wang XM, Ye JS, Lu NH, et al. MicroRNA-99a inhibits insulin-induced proliferation, migration, dedifferentiation, and rapamycin resistance of vascular smooth muscle cells by inhibiting insulin-like growth factor-1 receptor and mammalian target of rapamycin. Biochem Biophys Res Commun. 2017;486(2):414–22.  https://doi.org/10.1016/j.bbrc.2017.03.056.CrossRefPubMedGoogle Scholar
  26. 26.
    Tao J, Yang X, Han Z, Lu P, Wang J, Liu X, et al. Serum MicroRNA-99a helps detect acute rejection in renal transplantation. Transplant Proc. 2015;47(6):1683–7.  https://doi.org/10.1016/j.transproceed.2015.04.094.CrossRefPubMedGoogle Scholar
  27. 27.
    Yang SY, Wang YQ, Gao HM, Wang B, He Q. The clinical value of circulating miR-99a in plasma of patients with acute myocardial infarction. Eur Rev Med Pharmacol Sci. 2016;20(24):5193–7.PubMedGoogle Scholar
  28. 28.
    Jin Y, Tymen SD, Chen D, Fang ZJ, Zhao Y, Dragas D, et al. MicroRNA-99 family targets AKT/mTOR signaling pathway in dermal wound healing. PLoS One. 2013;8(5):e64434.  https://doi.org/10.1371/journal.pone.0064434.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Yao G, Yin M, Lian J, Tian H, Liu L, Li X, et al. MicroRNA-224 is involved in transforming growth factor-beta-mediated mouse granulosa cell proliferation and granulosa cell function by targeting Smad4. Mol Endocrinol. 2010;24(3):540–51.  https://doi.org/10.1210/me.2009-0432.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Lei L, Jin S, Gonzalez G, Behringer RR, Woodruff TK. The regulatory role of Dicer in folliculogenesis in mice. Mol Cell Endocrinol. 2010;315(1–2):63–73.  https://doi.org/10.1016/j.mce.2009.09.021.CrossRefPubMedGoogle Scholar
  31. 31.
    Hong X, Luense LJ, McGinnis LK, Nothnick WB, Christenson LK. Dicer1 is essential for female fertility and normal development of the female reproductive system. Endocrinology. 2008;149(12):6207–12.  https://doi.org/10.1210/en.2008-0294.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Ding CF, Chen WQ, Zhu YT, Bo YL, Hu HM, Zheng RH. Circulating microRNAs in patients with polycystic ovary syndrome. Hum Fertil. 2015;18(1):22–9.  https://doi.org/10.3109/14647273.2014.956811.CrossRefGoogle Scholar
  33. 33.
    Kota J, Chivukula RR, O'Donnell KA, Wentzel EA, Montgomery CL, Hwang HW, et al. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model. Cell. 2009;137(6):1005–17.  https://doi.org/10.1016/j.cell.2009.04.021.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Teixeira Filho FL, Baracat EC, Lee TH, Suh CS, Matsui M, Chang RJ, et al. Aberrant expression of growth differentiation factor-9 in oocytes of women with polycystic ovary syndrome. J Clin Endocrinol Metab. 2002;87(3):1337–44.  https://doi.org/10.1210/jcem.87.3.8316.CrossRefPubMedGoogle Scholar
  35. 35.
    Li D, Liu X, Lin L, Hou J, Li N, Wang C, et al. MicroRNA-99a inhibits hepatocellular carcinoma growth and correlates with prognosis of patients with hepatocellular carcinoma. J Biol Chem. 2011;286(42):36677–85.  https://doi.org/10.1074/jbc.M111.270561.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Brahmkhatri VP, Prasanna C, Atreya HS. Insulin-like growth factor system in cancer: novel targeted therapies. Biomed Res Int. 2015;2015:538019–24.  https://doi.org/10.1155/2015/538019.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Stubbs SA, Webber LJ, Stark J, Rice S, Margara R, Lavery S, et al. Role of insulin-like growth factors in initiation of follicle growth in normal and polycystic human ovaries. J Clin Endocrinol Metab. 2013;98(8):3298–305.  https://doi.org/10.1210/jc.2013-1378.CrossRefPubMedGoogle Scholar
  38. 38.
    Louhio H, Hovatta O, Sjoberg J, Tuuri T. The effects of insulin, and insulin-like growth factors I and II on human ovarian follicles in long-term culture. Mol Hum Reprod. 2000;6(8):694–8.CrossRefPubMedGoogle Scholar
  39. 39.
    Premoli AC, Santana LF, Ferriani RA, Moura MD, De Sa MF, Reis RM. Growth hormone secretion and insulin-like growth factor-1 are related to hyperandrogenism in nonobese patients with polycystic ovary syndrome. Fertil Steril. 2005;83(6):1852–5.  https://doi.org/10.1016/j.fertnstert.2004.10.057.CrossRefPubMedGoogle Scholar
  40. 40.
    Velazquez MA, Hermann D, Kues WA, Niemann H. Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor. Reproduction. 2011;141(1):91–103.  https://doi.org/10.1530/REP-10-0336.CrossRefPubMedGoogle Scholar
  41. 41.
    Luo L, Wang Q, Chen M, Yuan G, Wang Z, Zhou C. IGF-1 and IGFBP-1 in peripheral blood and decidua of early miscarriages with euploid embryos: comparison between women with and without PCOS. Gynecol Endocrinol. 2016;32(7):538–42.  https://doi.org/10.3109/09513590.2016.1138459.CrossRefPubMedGoogle Scholar
  42. 42.
    Homburg R, Pariente C, Lunenfeld B, Jacobs HS. The role of insulin-like growth factor-1 (IGF-1) and IGF binding protein-1 (IGFBP-1) in the pathogenesis of polycystic ovary syndrome. Hum Reprod. 1992;7(10):1379–83.CrossRefPubMedGoogle Scholar
  43. 43.
    Thierry van Dessel HJ, Lee PD, Faessen G, Fauser BC, Giudice LC. Elevated serum levels of free insulin-like growth factor I in polycystic ovary syndrome. J Clin Endocrinol Metab. 1999;84(9):3030–5.  https://doi.org/10.1210/jcem.84.9.5941.PubMedGoogle Scholar
  44. 44.
    Li W, Wang J, Chen QD, Qian X, Li Q, Yin Y, et al. Insulin promotes glucose consumption via regulation of miR-99a/mTOR/PKM2 pathway. PLoS One. 2013;8(6):e64924.  https://doi.org/10.1371/journal.pone.0064924.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Cai G, Ma X, Chen B, Huang Y, Liu S, Yang H, et al. MicroRNA-145 negatively regulates cell proliferation through targeting IRS1 in isolated ovarian granulosa cells from patients with polycystic ovary syndrome. Reprod Sci. 2016;24(6):902–10.  https://doi.org/10.1177/1933719116673197.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Reproductive Medicine Center, Tongji Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanPeople’s Republic of China
  2. 2.Department of Urology, Tongji Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanPeople’s Republic of China

Personalised recommendations