H19 gene methylation status is associated with male infertility

Li,Xiao‑Ping; Hao,Chao‑Liang; Wang,Qian; Yi,Xiao‑Mei; Jiang,Zhi‑Sheng

doi:10.3892/etm.2016.3314

H19 gene methylation status is associated with male infertility

Authors:
- Xiao‑Ping Li
- Chao‑Liang Hao
- Qian Wang
- Xiao‑Mei Yi
- Zhi‑Sheng Jiang
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Affiliations: Center of Reproductive Medicine, The First Affiliated Hospital, University of South China, Hengyang, Hunan 421000, P.R. China, Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan, University of South China, Hengyang, Hunan 421000, P.R. China
Published online on: May 9, 2016 https://doi.org/10.3892/etm.2016.3314
Pages: 451-456

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Abstract

The present study investigated the H19 gene methylation status in male infertility. Between March 2013 and June 2014, semen samples were collected from 15 normal fertile males and 15 males experiencing infertility, and routine analysis and sperm morphological assessment were performed. The semen samples were subjected to density gradient centrifugation to separate the sperm fraction, and genomic DNA from the sperms was extracted and treated for bisulfite modification. Following in vitro amplification by polymerase chain reaction (PCR), the purified PCR products were cloned into pMD®18‑T vectors and successful cloning was confirmed by restriction enzyme digestion. Positive clones were sequenced and the DNA methylation status was analyzed. The overall methylation rate in the normal fertile group was 100% (270/270), whereas in the infertile group the methylation rate was lower at 94.1% (525/558), revealing a statistically significant decrease in overall methylation rate in the infertile patients compared with the control group (χ2=15.12; P<0.001). The average methylation rates of CpG 1, 3 and 6 in the infertile group were statistically different from those in the normal control group (all P<0.05). The abnormal methylation of imprinted gene H19 is associated with male infertility, suggesting that H19 may serve as a biomarker for the detection of defects in human spermiogenesis.

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1	Kasturi SS, Tannir J and Brannigan RE: The metabolic syndrome and male infertility. J Androl. 29:251–259. 2008. View Article : Google Scholar : PubMed/NCBI
2	Jungwirth A, Giwercman A, Tournaye H, Diemer T, Kopa Z, Dohle G and Krausz C: European Association of Urology Working Group on Male Infertility: European association of urology guidelines on male infertility: The 2012 update. Eur Urol. 62:324–332. 2012. View Article : Google Scholar : PubMed/NCBI
3	Dada R, Kumar M, Jesudasan R, Fernández JL, Gosálvez J and Agarwal A: Epigenetics and its role in male infertility. J Assist Reprod Genet. 29:213–223. 2012. View Article : Google Scholar : PubMed/NCBI
4	Rajender S, Avery K and Agarwal A: Epigenetics, spermatogenesis and male infertility. Mutat Res. 727:62–71. 2011. View Article : Google Scholar : PubMed/NCBI
5	Wu W, Shen O, Qin Y, Lu J, Niu X, Zhou Z, Lu C, Xia Y, Wang S and Wang X: Methylenetetrahydrofolate reductase C677T polymorphism and the risk of male infertility: A meta-analysis. Int J Androl. 35:18–24. 2012. View Article : Google Scholar : PubMed/NCBI
6	Ferlin A, Vinanzi C, Selice R, Garolla A, Frigo AC and Foresta C: Toward a pharmacogenetic approach to male infertility: Polymorphism of follicle-stimulating hormone beta-subunit promoter. Fertil Steril. 96:1344–1349.e2. 2011. View Article : Google Scholar : PubMed/NCBI
7	Hammoud AO, Meikle AW, Reis LO, Gibson M, Peterson CM and Carrell DT: Obesity and male infertility: A practical approach. Semin Reprod Med. 30:486–495. 2012. View Article : Google Scholar : PubMed/NCBI
8	Ross C, Morriss A, Khairy M, Khalaf Y, Braude P, Coomarasamy A and El-Toukhy T: A systematic review of the effect of oral antioxidants on male infertility. Reprod Biomed Online. 20:711–723. 2010. View Article : Google Scholar : PubMed/NCBI
9	Ferlin A, Raicu F, Gatta V, Zuccarello D, Palka G and Foresta C: Male infertility: Role of genetic background. Reprod Biomed Online. 14:734–745. 2007. View Article : Google Scholar : PubMed/NCBI
10	Poongothai J, Gopenath TS and Manonayaki S: Genetics of human male infertility. Singapore Med J. 50:336–347. 2009.PubMed/NCBI
11	Krausz C and Giachini C: Genetic risk factors in male infertility. Arch Androl. 53:125–133. 2007. View Article : Google Scholar : PubMed/NCBI
12	Gabory A, Ripoche MA, Le Digarcher A, Watrin F, Ziyyat A, Forné T, Jammes H, Ainscough JF, Surani MA, Journot L and Dandolo L: H19 acts as a trans regulator of the imprinted gene network controlling growth in mice. Development. 136:3413–3421. 2009. View Article : Google Scholar : PubMed/NCBI
13	Zhang Y and Tycko B: Monoallelic expression of the human H19 gene. Nat Genet. 1:40–44. 1992. View Article : Google Scholar : PubMed/NCBI
14	Bartolomei MS, Zemel S and Tilghman SM: Parental imprinting of the mouse H19 gene. Nature. 351:153–155. 1991. View Article : Google Scholar : PubMed/NCBI
15	Reese KJ, Lin S, Verona RI, Schultz RM and Bartolomei MS: Maintenance of paternal methylation and repression of the imprinted H19 gene requires MBD3. PLoS Genet. 3:e1372007. View Article : Google Scholar : PubMed/NCBI
16	Guttman M and Rinn JL: Modular regulatory principles of large non-coding RNAs. Nature. 482:339–346. 2012. View Article : Google Scholar : PubMed/NCBI
17	Cai X and Cullen BR: The imprinted H19 noncoding RNA is a primary microRNA precursor. RNA. 13:313–316. 2007. View Article : Google Scholar : PubMed/NCBI
18	Matouk IJ, DeGroot N, Mezan S, Ayesh S, Abu-lail R, Hochberg A and Galun E: The H19 non-coding RNA is essential for human tumor growth. PLoS One. 2:e8452007. View Article : Google Scholar : PubMed/NCBI
19	Steck E, Boeuf S, Gabler J, Werth N, Schnatzer P, Diederichs S and Richter W: Regulation of H19 and its encoded microRNA-675 in osteoarthritis and under anabolic and catabolic in vitro conditions. J Mol Med (Berl). 90:1185–1195. 2012. View Article : Google Scholar : PubMed/NCBI
20	Tsang WP, Ng EK, Ng SS, Jin H, Yu J, Sung JJ and Kwok TT: Oncofetal H19-derived miR-675 regulates tumor suppressor RB in human colorectal cancer. Carcinogenesis. 31:350–358. 2010. View Article : Google Scholar : PubMed/NCBI
21	Boissonnas CC, Abdalaoui HE, Haelewyn V, Fauque P, Dupont JM, Gut I, Vaiman D, Jouannet P, Tost J and Jammes H: Specific epigenetic alterations of IGF2-H19 locus in spermatozoa from infertile men. Eur J Hum Genet. 18:73–80. 2010. View Article : Google Scholar : PubMed/NCBI
22	Marques CJ, Costa P, Vaz B, Carvalho F, Fernandes S, Barros A and Sousa M: Abnormal methylation of imprinted genes in human sperm is associated with oligozoospermia. Mol Hum Reprod. 14:67–74. 2008. View Article : Google Scholar : PubMed/NCBI
23	Macklin R: Revising the Declaration of Helsinki: A work in progress. Indian J Med Ethics. 9:224–226. 2012.PubMed/NCBI
24	World Health Organization: Laboratory manual of the WHO for the examination of human semen and sperm-cervical mucus interaction. Ann Ist Super Sanita. 37:I–XII. 2001.(In Italian). PubMed/NCBI
25	Ferguson-Smith AC: Genomic imprinting: The emergence of an epigenetic paradigm. Nat Rev Genet. 12:565–575. 2011. View Article : Google Scholar : PubMed/NCBI
26	Kobayashi H, Sato A, Otsu E, Hiura H, Tomatsu C, Utsunomiya T, Sasaki H, Yaegashi N and Arima T: Aberrant DNA methylation of imprinted loci in sperm from oligospermic patients. Hum Mol Genet. 16:2542–2551. 2007. View Article : Google Scholar : PubMed/NCBI
27	Kato Y and Nozaki M: Distinct DNA methylation dynamics of spermatogenic cell-specific intronless genes is associated with CpG content. PLoS One. 7:e436582012. View Article : Google Scholar : PubMed/NCBI
28	Dammann RH, Kirsch S, Schagdarsurengin U, Dansranjavin T, Gradhand E, Schmitt WD and Hauptmann S: Frequent aberrant methylation of the imprinted IGF2/H19 locus and LINE1 hypomethylation in ovarian carcinoma. Int J Oncol. 36:171–179. 2010.PubMed/NCBI
29	Poplinski A, Tuttelmann F, Kanber D, Horsthemke B and Gromoll J: Idiopathic male infertility is strongly associated with aberrant methylation of MEST and IGF2/H19 ICR1. Int J Androl. 33:642–649. 2010.PubMed/NCBI