Hsa-miR-27a-3p overexpression in men with nonobstructive azoospermia: A case-control study


Background: The role of KDM3A and its downstream genes in male fertility has been approved in animal models. Additionally, the expression shrinkage of KDM3A is significantly correlated with human azoospermia phenotype. Aberrant expression of micro-RNAs could mislead spermatogenesis and mostly lead to diverse phenotypes of male infertility.

Objective: The aim of this study was to evaluate the expression level of hsa-miR-27a- 3p in azoospermic men to reveal its possible association with infertility.

Materials and Methods: This case-control study was conducted on 30 azoospermic men, of whom, 19 had non obstructive azoospermia (NOA) and 11 obstructive azoospermia (OA) according to the pathological examinations. Comprehensive bioinformatics investigations were performed securely and hsa-miR-27a-3p was selected afterward. Reverse Transcriptase-quantitative polymerase chain reaction (RTqPCR) method was used and statistical analysis was performed to compare the expression level of hsa-miR-27a-3p in both OA and NOA individuals.

Results: In silico analysis suggested hsa-miR-27a-3p, with its potential binding ability to target KDM3A transcripts. The expression analysis of candidate hsa-miR-27a-3p indicated its significant overexpression in NOA men.

Conclusion: The hsa-miR-27a-3p was overexpressed in NOA men compared to OA-control individuals. As a consequence, the overexpressed micro-RNA could downregulate directly KDM3A and indirectly TNP1 and PRM1. Therefore, spermatogenesis could be misled and male infertility could be developed.

Key words: hsa-miR-27a-3p, Male infertility, KDM3A.

[1] Fernández Pelegrina R, Kessler AG, Rawlins RG. Modern approaches to the treatment of human infertility through assisted reproduction. P R Health Sci J 1991; 10: 75–81.

[2] Cooper TG, Noonan E, Von Eckardstein S, Auger J, Baker HW, Behre HM, et al. World Health Organization reference values for human semen characteristics. Hum Reprod Update 2010; 16: 231– 245.

[3] Irvine DS. Epidemiology and aetiology of male infertility. Hum Reprod 1998; 13 (Suppl.): 33–44.

[4] Aziz N. The importance of semen analysis in the context of azoospermia. Clinics 2013; 68 (Suppl.): 35–38.

[5] Jarow JP, Espeland MA, Lipshultz LI. Evaluation of the azoospermic patient. J Urol 1989; 142: 62–65.

[6] Okada Y, Tateishi K, Zhang Y. Histone demethylase JHDM2A is involved in male infertility and obesity. J Androl 2010; 31: 75–78.

[7] Javadirad SM, Hojati Z, Ghaedi K, Nasr−Esfahani MH. Expression ratio of histone demethylase KDM 3A to protamine−1 mRNA is predictive of successful testicular sperm extraction in men with obstructive and non−obstructive azoospermia. Andrology 2016; 4: 492–499.

[8] Okada Y, Scott G, Ray MK, Mishina Y, Zhang Y. Histone demethylase JHDM2A is critical for Tnp1 and Prm1 transcription and spermatogenesis. Nature 2007; 450: 119–123.

[9] Zamudio NM, Chong S, O’Bryan MK. Epigenetic regulation in male germ cells. Reproduction 2008; 136: 131–146.

[10] Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 2010; 11: 597–610.

[11] Heneghan HM, Miller N, Kerin MJ. MiRNAs as biomarkers and therapeutic targets in cancer. Curr Opin Pharmacol 2010; 10: 543–550.

[12] Mattick JS. Non−coding RNAs: the architects of eukaryotic complexity. EMBO Rep 2001; 2: 986–991.

[13] Charan J, Biswas T. How to calculate sample size for different study designs in medical research? Indian J Psychol Med 2013; 35: 121–126.

[14] Bonaparte E, Moretti M, Colpi GM, Nerva F, Contalbi G, Vaccalluzzo L, et al. ESX1 gene expression as a robust marker of residual spermatogenesis in azoospermic men. Hum Reprod 2010; 25: 1398– 1403.

[15] Pansa A, Sirchia SM, Melis S, Giacchetta D, Castiglioni M, Colapietro P, et al. ESX1 mRNA expression in seminal fluid is an indicator of residual spermatogenesis in non-obstructive azoospermic men. Hum Reprod 2014; 29: 2620–2627.

[16] Schlegel PN, Li PS. Microdissection TESE: sperm retrieval in non-obstructive azoopsermia. Hum Reprod Update 1998; 4: 439.

[17] Song GJ, Lee H, Park Y, Lee HJ, Lee YS, Seo JT, et al. Expression pattern of germ cell-specific genes in the testis of patients with nonobstructive azoospermia: usefulness as a molecular marker to predict the presence of testicular sperm. Fertil Steril 2000; 73: 1104–1108.

[18] Seo JT, Ko WJ. Predictive factors of successful testicular sperm recovery in non−obstructive azoospermia patients. Int J Androl 2001; 24: 306– 310.

[19] Tournaye H, Verheyen G, Nagy P, Ubaldi F, Goossens A, Silber S, et al. Are there any predictive factors for successful testicular sperm recovery in azoospermic patients? Hum Reprod 1997; 12: 80–86.

[20] Brugo-Olmedo S, De Vincentiis S, Calamera JC, Urrutia F, Nodar F, Acosta AA. Serum inhibin B may be a reliable marker of the presence of testicular spermatozoa in patients with nonobstructive azoospermia. Fertil Steril 2001; 76: 1124–1129.

[21] Meachem SJ, Nieschlag E, Simoni M. Inhibin B in male reproduction: pathophysiology and clinical relevance. Eur J Endocrinol 2001; 145: 561–571.

[22] Lee JH, Lee DR, Yoon SJ, Chai YG, Roh SI, Yoon HS. Expression of DAZ (deleted in azoospermia), DAZL1 (DAZ-like) and protamine-2 in testis and its application for diagnosis of spermatogenesis in nonobstructive azoospermia. Mol Hum Reprod 1998; 4: 827–834.

[23] Kleiman SE, Yogev L, Hauser R, Botchan A, Maymon BS, Paz G, et al. Expression profile of AZF genes in testicular biopsies of azoospermic men. Hum Reprod 2007; 22: 151–158.

[24] Wu W, Qin Y, Li Z, Dong J, Dai J, Lu C, et al. Genome-wide microRNA expression profiling in idiopathic non-obstructive azoospermia: significant up-regulation of miR-141, miR-429 and miR-7-1-3p. Hum Reprod 2013; 28: 1827–1836.

[25] Abu-Halima M, Hammadeh M, Schmitt J, Leidinger P, Keller A, Meese E, et al. Altered microRNA expression profiles of human spermatozoa in patients with different spermatogenic impairments. Fertil Steril 2013; 99: 1249–1255. e16.

[26] Zhuang X, Li Z, Lin H, Gu L, Lin Q, Lu Z, et al. Integrated miRNA and mRNA expression profiling to identify mRNA targets of dysregulated miRNAs in non-obstructive azoospermia. Sci Rep 2015; 5: 7922.

[27] Cui Z, Liu G, Kong D. miRNA-27a promotes the proliferation and inhibits apoptosis of human pancreatic cancer cells by Wnt/β-catenin pathway. Oncol Rep 2018; 39: 755–763.

[28] Fang F, Huang B, Sun S, Xiao M, Guo J, Yi X, et al. miR-27a inhibits cervical adenocarcinoma progression by downregulating the TGF-βRI signaling pathway. Cell Death Dis 2018; 9: 395–408.

[29] Gao W, Hong Z, Huang H, Zhu A, Lin S, Cheng C, et al. miR-27a in serum acts as biomarker for prostate cancer detection and promotes cell proliferation by targeting Sprouty2. Oncol Lett 2018; 16: 5291–5298.

[30] Wang YL, Gong WG, Yuan QL. Effects of miR- 27a upregulation on thyroid cancer cells migration, invasion, and angiogenesis. Genet Mol Res 2016; 15: 1–10.