Archives of Gynecology and Obstetrics

, Volume 299, Issue 6, pp 1715–1725 | Cite as

Novel circular RNA expression in the cumulus cells of patients with polycystic ovary syndrome

  • Zhi Ma
  • Huishan Zhao
  • Yan Zhang
  • Xiaoyan Liu
  • Cuifang HaoEmail author
Gynecologic Endocrinology and Reproductive Medicine



Circular RNAs (circRNAs) mediate the posttranscriptional regulation of multiple genes by functioning as microRNA (miRNA) sponges. This study aimed to detect the novel expression of circRNAs in the cumulus cells (CCs) of polycystic ovary syndrome (PCOS) patients and their potential significance in the pathogenesis of PCOS.


circRNAs in the CCs from 6 PCOS patients and 6 normal control individuals were collected for microarray analysis, and an independent cohort study including 25 PCOS patients and 25 normal control individuals was conducted to validate the circRNA microarray results using quantitative real-time polymerase chain reaction (qRT-PCR). Spearman’s rank correlation and receiver operating characteristic (ROC) were performed to delineate the potential correlation between novel circRNAs and patients’ clinical characteristics and their potential efficacy for the diagnosis of PCOS. Bioinformatics analysis was applied to investigate the potential roles of circRNAs in the pathogenesis of PCOS.


A total of 286 circRNAs (167 upregulated and 119 downregulated) were identified by microarray that was differentially expressed between the PCOS and non-PCOS groups. After qRT-PCR validation, the expression levels of hsa_circ_0043533 (p < 0.05) and hsa_circ_0043532 (p < 0.01) were significantly higher in the PCOS group, while the expression level of hsa_circ_0097636 (p < 0.01) was prominently lower versus the non-PCOS group. Spearman’s rank correlation indicated that the serum testosterone (T) level positively correlated with the expression of hsa_circ_0043533 and hsa_circ_0097636 in the PCOS group. The ROC curve analysis found that the combination of hsa_circ_0097636 and T resulted in a larger area under the curve (AUC) (0.893) compared with each circRNA alone (0.709, 0.738, and 0.718 for hsa_circ_0043533, hsa_circ_0097636 and hsa_circ_0043532, respectively). Bioinformatics analysis revealed that the dysregulated circRNAs were potentially involved in cell cycle, oocyte meiosis, progesterone-mediated oocyte maturation, the FOXO signaling pathway, phosphatidylinositol signaling and glycerophospholipid metabolism.


The expression of circRNAs in CCs was significantly different between PCOS and normal control individuals. We validated three circRNAs, which could lead to a better understanding of disease pathogenesis and the development of effective therapeutic interventions for PCOS patients.


Polycystic ovary syndrome Cumulus cells Circular RNA MicroRNA sponge Posttranscriptional regulation 



This work was financially supported by the National Natural Science Foundation of China (No. 81671416).

Author contributions

ZM: data collection, formal analysis, manuscript writing. HZ: methodology and software supporting. YZ: methodology supporting and manuscript editing. XL: sample collection. CH: project development and manuscript editing.

Compliance with ethical standards

Conflict of interest

We declare no conflict of interest.

Ethical statement

This study was approved by the Institutional Ethics Review Board, Yantai Yuhuangding Hospital affiliated to Qingdao University.

Supplementary material

404_2019_5122_MOESM1_ESM.xlsx (15 kb)
Supplementary material 1 (XLSX 15 kb)


  1. 1.
    Yildiz BO, Bozdag G, Yapici Z et al (2012) Prevalence, phenotype and cardiometabolic risk of polycystic ovary syndrome under different diagnostic criteria. Hum Reprod 27:3067–3073CrossRefGoogle Scholar
  2. 2.
    Broekmans FJ, Knauff EA, Valkenburg O et al (2006) PCOS according to the Rotterdam consensus criteria: change in prevalence among WHO-II anovulation and association with metabolic factors. BJOG 113:1210–1217CrossRefGoogle Scholar
  3. 3.
    Diao FY, Xu M, Hu Y et al (2004) The molecular characteristics of polycystic ovary syndrome (PCOS) ovary defined by human ovary cDNA microarray. J Mol Endocrinol 33:59–72CrossRefGoogle Scholar
  4. 4.
    Teede HJ, Misso ML, Deeks AA et al (2011) Assessment and management of polycystic ovary syndrome: summary of an evidence-based guideline. Med J Aust 195:S65–112CrossRefGoogle Scholar
  5. 5.
    Salzman J (2016) Circular RNA expression: its potential regulation and function. Trends Genet 32:309–316CrossRefGoogle Scholar
  6. 6.
    Qu S, Yang X, Li X et al (2015) Circular RNA: a new star of noncoding RNAs. Cancer Lett 365:141–148CrossRefGoogle Scholar
  7. 7.
    Rybak-Wolf A, Stottmeister C, Glazar P et al (2015) Circular RNAs in the mammalian brain are highly abundant, conserved, and dynamically expressed. Mol Cell 58:870–885CrossRefGoogle Scholar
  8. 8.
    Legnini I, Di Timoteo G, Rossi F et al (2017) Circ-ZNF609 is a circular RNA that can be translated and functions in myogenesis. Mol Cell 66:22–37.e29CrossRefGoogle Scholar
  9. 9.
    Shan K, Liu C, Liu BH et al (2017) Circular noncoding RNA HIPK3 mediates retinal vascular dysfunction in diabetes mellitus. Circulation 136:1629–1642CrossRefGoogle Scholar
  10. 10.
    Jeck WR, Sharpless NE (2014) Detecting and characterizing circular RNAs. Nat Biotechnol 32:453–461CrossRefGoogle Scholar
  11. 11.
    Dang Y, Yan L, Hu B et al (2016) Tracing the expression of circular RNAs in human pre-implantation embryos. Genome Biol 17:130CrossRefGoogle Scholar
  12. 12.
    Cheng J, Huang J, Yuan S et al (2017) Circular RNA expression profiling of human granulosa cells during maternal aging reveals novel transcripts associated with assisted reproductive technology outcomes. PLoS One 12:e0177888CrossRefGoogle Scholar
  13. 13.
    Rotterdam EA-SPCWG (2004) Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril 81:19–25Google Scholar
  14. 14.
    Huang X, Liu C, Hao C et al (2016) Identification of altered microRNAs and mRNAs in the cumulus cells of PCOS patients: miRNA-509-3p promotes oestradiol secretion by targeting MAP3 K8. Reproduction 151:643–655CrossRefGoogle Scholar
  15. 15.
    You X, Vlatkovic I, Babic A et al (2015) Neural circular RNAs are derived from synaptic genes and regulated by development and plasticity. Nat Neurosci 18:603–610CrossRefGoogle Scholar
  16. 16.
    Ashwal-Fluss R, Meyer M, Pamudurti NR et al (2014) circRNA biogenesis competes with pre-mRNA splicing. Mol Cell 56:55–66CrossRefGoogle Scholar
  17. 17.
    Zhang Y, Zhang XO, Chen T et al (2013) Circular intronic long noncoding RNAs. Mol Cell 51:792–806CrossRefGoogle Scholar
  18. 18.
    Li Z, Huang C, Bao C et al (2015) Exon-intron circular RNAs regulate transcription in the nucleus. Nat Struct Mol Biol 22:256–264CrossRefGoogle Scholar
  19. 19.
    Yang JL, Zhang CP, Li L et al (2010) Testosterone induces redistribution of forkhead box-3a and down-regulation of growth and differentiation factor 9 messenger ribonucleic acid expression at early stage of mouse folliculogenesis. Endocrinology 151:774–782CrossRefGoogle Scholar
  20. 20.
    Chen YX, Zhang XJ, Huang J et al (2016) UHPLC/Q-TOFMS-based plasma metabolomics of polycystic ovary syndrome patients with and without insulin resistance. J Pharm Biomed Anal 121:141–150CrossRefGoogle Scholar
  21. 21.
    Rosenfield RL, Ehrmann DA (2016) The pathogenesis of polycystic ovary syndrome (PCOS): the hypothesis of PCOS as functional ovarian hyperandrogenism revisited. Endocr Rev 37:467–520CrossRefGoogle Scholar
  22. 22.
    Chazenbalk G, Chen YH, Heneidi S et al (2012) Abnormal expression of genes involved in inflammation, lipid metabolism, and Wnt signaling in the adipose tissue of polycystic ovary syndrome. J Clin Endocrinol Metab 97:E765–770CrossRefGoogle Scholar
  23. 23.
    Zhao Y, Zhang C, Huang Y et al (2015) Up-regulated expression of WNT5a increases inflammation and oxidative stress via PI3K/AKT/NF-kappaB signaling in the granulosa cells of PCOS patients. J Clin Endocrinol Metab 100:201–211CrossRefGoogle Scholar
  24. 24.
    Barrett SP, Salzman J (2016) Circular RNAs: analysis, expression and potential functions. Development 143:1838–1847CrossRefGoogle Scholar
  25. 25.
    Memczak S, Jens M, Elefsinioti A et al (2013) Circular RNAs are a large class of animal RNAs with regulatory potency. Nature 495:333–338CrossRefGoogle Scholar
  26. 26.
    Wang K, Long B, Liu F et al (2016) A circular RNA protects the heart from pathological hypertrophy and heart failure by targeting miR-223. Eur Heart J 37:2602–2611CrossRefGoogle Scholar
  27. 27.
    Piwecka M, Glazar P, Hernandez-Miranda LR et al (2017) Loss of a mammalian circular RNA locus causes miRNA deregulation and affects brain function. Science 357:6357CrossRefGoogle Scholar
  28. 28.
    Chen YH, Heneidi S, Lee JM et al (2013) miRNA-93 inhibits GLUT4 and is overexpressed in adipose tissue of polycystic ovary syndrome patients and women with insulin resistance. Diabetes 62:2278–2286CrossRefGoogle Scholar
  29. 29.
    Cai G, Ma X, Chen B et al (2017) MicroRNA-145 negatively regulates cell proliferation through targeting IRS1 in isolated ovarian granulosa cells from patients with polycystic ovary syndrome. Cell Biochem Funct 24:902–910Google Scholar
  30. 30.
    Zhang CL, Wang H, Yan CY et al (2017) Deregulation of RUNX2 by miR-320a deficiency impairs steroidogenesis in cumulus granulosa cells from polycystic ovary syndrome (PCOS) patients. Biochem Biophys Res Commun 482:1469–1476CrossRefGoogle Scholar
  31. 31.
    Sen A, Prizant H, Light A et al (2014) Androgens regulate ovarian follicular development by increasing follicle stimulating hormone receptor and microRNA-125b expression. Proc Natl Acad Sci USA 111:3008–3013CrossRefGoogle Scholar
  32. 32.
    Long W, Zhao C, Ji C et al (2014) Characterization of serum microRNAs profile of PCOS and identification of novel non-invasive biomarkers. Cell Physiol Biochem 33:1304–1315CrossRefGoogle Scholar
  33. 33.
    Sang Q, Yao Z, Wang H et al (2013) Identification of microRNAs in human follicular fluid: characterization of microRNAs that govern steroidogenesis in vitro and are associated with polycystic ovary syndrome in vivo. J Clin Endocrinol Metab 98:3068–3079CrossRefGoogle Scholar
  34. 34.
    Sirotkin AV, Ovcharenko D, Grossmann R et al (2009) Identification of microRNAs controlling human ovarian cell steroidogenesis via a genome-scale screen. J Cell Physiol 219:415–420CrossRefGoogle Scholar
  35. 35.
    Donadeu FX, Schauer SN, Sontakke SD (2012) Involvement of miRNAs in ovarian follicular and luteal development. J Endocrinol 215:323–334CrossRefGoogle Scholar
  36. 36.
    Yao N, Yang BQ, Liu Y et al (2010) Follicle-stimulating hormone regulation of microRNA expression on progesterone production in cultured rat granulosa cells. Endocrine 38:158–166CrossRefGoogle Scholar
  37. 37.
    Sathyapalan T, David R, Gooderham NJ et al (2015) Increased expression of circulating miRNA 93 in women with polycystic ovary syndrome may represent a novel, non-invasive biomarker for diagnosis. Sci Rep 5:16890CrossRefGoogle Scholar
  38. 38.
    Scalici E, Traver S, Mullet T et al (2016) Circulating microRNAs in follicular fluid, powerful tools to explore in vitro fertilization process. Sci Rep 6(24976):3Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Medical College of Qingdao UniversityQingdaoPeople’s Republic of China
  2. 2.Centre for Reproductive MedicineYantai Yuhuangding Hospital Affiliated to Qingdao UniversityYantaiPeople’s Republic of China
  3. 3.Institute of ZoologyChinese Academy of SciencesBeijingPeople’s Republic of China

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