Significance of microRNA targeted estrogen receptor in male fertility

Document Type: Original Article


1 Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran Women’s Reproductive Health Research Center, Alzahra Hospital, Tabriz, Iran

2 Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

3 Women’s Reproductive Health Research Center, Alzahra Hospital, Tabriz, Iran

4 Research Institute for Fundamental Sciences (RIFS), University of Tabriz, Tabriz, Iran

5 Department of Biotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical Sciences, Tabriz, Iran


Objective(s): Estrogen receptor-alpha (ERα) mediates estrogen action in regulation of different levels of the hypothalamic-pituitary-testis axis. It has a key role in spermatogenesis. Estrogen receptor alpha knock-out (ER koα) male mice were infertile and severe impairment in spermatogenesis and seminiferous tubules was observed. Recently, it has been reported that microRNA (miRNA) mir-100 and let-7b were predicted to target ERα gene. MiRNA are small, endogenous, single stranded RNA molecules that regulate gene expression and have been implicated in various disease states. It has been proved that some miRNAs expression is tissue- and disease-specific, giving potential for identifying miRNAs as a diagnostic tool.
Materials and Methods: In this study, the change in the expression levels of mir-100, let-7b and ERα expression levels were evaluated in oligospermic infertile patients (n=43) compared to control fertile subjects (n=43). After washing and separating sperms, total RNA was isolated and then cDNA was synthesized. The expression levels of mir-100 and let-7b and ERα were evaluated by real time PCR.
Results: Mir-100, let-7b levels were significantly higher than those in control group (P=0.008 and P=0.009, respectively). We have found that, ERα level was significantly decreased in comparison with normal group (P< 0.0001).
Conclusion: Changes in mir-100, let-7b and ERα expression levels in oligospermic patients may be associated with the susceptibility and progression of infertility. The results of this study indicate that miRNA can have a key role in spermatogenesis and might have a diagnostic and prognostic value in men infertility.


1. Definitions of infertility and recurrent pregnancy loss: a committee opinion. Fertil Steril 2013; 99:63.

2. Ford WC. Comments on the release of the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen. Asian J Androl 2010; 12:59-63.

3. Dohle GR, Colpi GM, Hargreave TB, Papp GK, Jungwirth A, Weidner W. EAU guidelines on male infertility. Eur Urol 2005; 48:703-711.

4. Dejarnette JM. The effect of semen quality on reproductive efficiency. Vet Clin North Am Food Anim Pract 2005;21:409-418.

5. Slezak R, Sasiadek M. [Chromosome Y microdeletions in the pathogenesis of male infertility]. Pol Merkur Lekarski 2002; 13:229-233.

6. Guo T, Qin Y, Gao X, Chen H, Li G, Ma J, et al. The role of male chromosomal polymorphism played in spermatogenesis and the outcome of IVF/ICSI-ET treatment. Int J Androl 2012; 35:802-809.

7. Carreau S, Silandre D, Bois C, Bouraima H, Galeraud-Denis I, Delalande C. Estrogens: a new player in spermatogenesis. Folia histochemica et cytobiologica 2007;45:5-10.

8. Ellmann S, Sticht H, Thiel F, Beckmann MW, Strick R, Strissel PL. Estrogen and progesterone receptors: from molecular structures to clinical targets. Cell Mol Life Sci 2009; 66:2405-2426.

9. Carreau S, Genissel C, Bilinska B, Levallet J. Sources of oestrogen in the testis and reproductive tract of the male. International journal of andrology. 1999;22:211-223.

10. Zhou J, Song T, Gong S, Zhong M, Su G. microRNA regulation of the expression of the estrogen receptor in endometrial cancer. Mol Med Rep 2010; 3:387-392.

11. Zhao Y, Deng C, Lu W, Xiao J, Ma D, Guo M, et al. let-7 microRNAs induce tamoxifen sensitivity by downregulation of estrogen receptor alpha signaling in breast cancer. Mol Med 2011; 17:1233-1241.

12. Ostermeier GC, Goodrich RJ, Moldenhauer JS, Diamond MP, Krawetz SA. A suite of novel human spermatozoal RNAs. J Androl 2005; 26:70-74.

13. He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 2004; 5:522-531.

14. Alvarez-Garcia I, Miska EA. MicroRNA functions in animal development and human disease. Development  2005; 132:4653-4662.

15. Ro S, Park C, Sanders KM, McCarrey JR, Yan W. Cloning and expression profiling of testis-expressed microRNAs. Dev Biol 2007; 311:592-602.

16. Huszar JM, Payne CJ. MicroRNA 146 (Mir146) modulates spermatogonial differentiation by retinoic acid in mice. Biol Rep 2013; 88:15.

17. Goodrich R, Johnson G, Krawetz SA. The preparation of human spermatozoal RNA for clinical analysis. Arch Androl 2007; 53:161-167.

18. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucl Acids Res 2001; 29:e45.

19. Yan W. Male infertility caused by spermiogenic defects: lessons from gene knockouts. Mol Cell Endocrinol 2009; 306:24-32.

20. Heidari MM, Khatami M, Talebi AR. The POLG Gene polymorphism in Iranian varicocele-associated infertility patients. Iran J Basic Med Sci 2012; 15:739-744.

21. Yadav RP, Kotaja N. Small RNAs in spermatogenesis. Mol Cell Endocrinol 2013; 28.

22. Lian J, Zhang X, Tian H, Liang N, Wang Y, Liang C, et al. Altered microRNA expression in patients with non-obstructive azoospermia. Reprod Biol Endocrinol 2009; 7:13.

23. Zhou J, Shen K, Zeng JF, Yang JX, Cao DY, Cui QC. [Differential expression of microRNAs associated with estrogen receptor alpha and progesterone receptor in typeIand typeII endometrial adenocarcinomas. Zhonghua Fu Chan ke Za Zhi 2009; 44:765-770.

24. Xu D, Tan J, Zhou M, Jiang B, Xie H, Nie X, et al. Let-7b and microRNA-199a inhibit the proliferation of B16F10 melanoma cells. Oncol Lett 2012; 4:941-946.

25. Guarducci E, Nuti F, Becherini L, Rotondi M, Balercia G, Forti G, et al. Estrogen receptor alpha promoter polymorphism: stronger estrogen action is coupled with lower sperm count. Hum Rep 2006; 21:994-1001.

26. Hess RA, Fernandes SA, Gomes GR, Oliveira CA, Lazari MF, Porto CS. Estrogen and its receptors in efferent ductules and epididymis. J Androl 2011; 32:600-613.

27. Gunawan A, Kaewmala K, Uddin MJ, Cinar MU, Tesfaye D, Phatsara C, et al. Association study and expression analysis of porcine ESR1 as a candidate gene for boar fertility and sperm quality. Anim Reprod Sci 2011; 128:11-21.

28. Joseph A, Shur BD, Ko C, Chambon P, Hess RA. Epididymal hypo-osmolality induces abnormal sperm morphology and function in the estrogen receptor alpha knockout mouse. Biol Reprod 2010; 82:958-967.

29. Guido C, Perrotta I, Panza S, Middea E, Avena P, Santoro M, et al. Human sperm physiology: estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta) influence sperm metabolism and may be involved in the pathophysiology of varicocele-associated male infertility. J Cell Physiol 2011; 226:3403-3412.

30. Safarinejad MR, Shafiei N, Safarinejad S. Association of polymorphisms in the estrogen receptors alpha, and beta (ESR1, ESR2) with the occurrence of male infertility and semen parameters. J Steroid Biochem Mol Biol 2010; 122:193-203.

31. Vigil P, Toro A, Godoy A. Physiological action of oestradiol on the acrosome reaction in human spermatozoa. Andrologia 2008; 40:146-151.

32. Hartmann M. Acrosome reaction and fertility. Int J Androl 1995; 18:53-55.

33. Liu DY, Baker HW. High frequency of defective sperm-zona pellucida interaction in oligozoospermic infertile men. Hum Reprod 2004; 19:228-233.

34. Liu de Y, Liu ML, Garrett C, Baker HW. Comparison of the frequency of defective sperm-zona pellucida (ZP) binding and the ZP-induced acrosome reaction between subfertile men with normal and abnormal semen. Hum Reprod 2007; 22:1878-1884.

35. Braydich-Stolle L, Kostereva N, Dym M, Hofmann MC. Role of Src family kinases and N-Myc in spermatogonial stem cell proliferation. Dev Biol 2007; 304:34-45.

36. Stabile V, Russo M, Chieffi P. 17beta-estradiol induces Akt-1 through estrogen receptor-beta in the frog (Rana esculenta) male germ cells. Reproduction. 2006; 132:477-484.

37. Kim ST, Omurtag K, Moley KH. Decreased spermatogenesis, fertility, and altered Slc2A expression in Akt1-/- and Akt2-/- testes and sperm. Reprod Sci 2012; 19:31-42.

38. McIver SC, Stanger SJ, Santarelli DM, Roman SD, Nixon B, McLaughlin EA. A unique combination of male germ cell miRNAs coordinates gonocyte differentiation. PloS one 2012; 7:e35553.

39. Poplinski A, Tuttelmann F, Kanber D, Horsthemke B, Gromoll J. Idiopathic male infertility is strongly associated with aberrant methylation of MEST and IGF2/H19 ICR1. Int J Androl 2010; 33:642-649.

40. Kerjean A, Dupont JM, Vasseur C, Le Tessier D, Cuisset L, Paldi A, et al. Establishment of the paternal methylation imprint of the human H19 and MEST/PEG1 genes during spermatogenesis. Hum Mol Genet 2000; 9:2183-2187.