title: H19gene methylation status is associated with male infertility.
abstract: The present study investigated theH19gene 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. Followingin vitroamplification 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 geneH19is associated with male infertility, suggesting thatH19may serve as a biomarker for the detection of defects in human spermiogenesis.
authors.affiliation: Center of Reproductive Medicine, The First Affiliated Hospital, University of South China, Hengyang, Hunan 421000, P.R. China; Post-Doctoral Mobile Stations for Basic Medicine, 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.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Li
authors.affiliation: Center of Reproductive Medicine, The First Affiliated Hospital, University of South China, Hengyang, Hunan 421000, P.R. China.
authors.forename: Chao-Liang
authors.initials: CL
authors.lastname: Hao
authors.affiliation: Center of Reproductive Medicine, The First Affiliated Hospital, University of South China, Hengyang, Hunan 421000, P.R. China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Center of Reproductive Medicine, The First Affiliated Hospital, University of South China, Hengyang, Hunan 421000, P.R. China.
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: Yi
authors.affiliation: Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan, University of South China, Hengyang, Hunan 421000, P.R. China.
authors.forename: Zhi-Sheng
authors.initials: ZS
authors.lastname: Jiang
keywords: male infertility; methylation; polymerase chain reaction; sperm morphology; the human H19 gene; vector clone