Alkhaled Y, Laqqan M, Tierling S, Lo Porto C, Amor H, Hammadeh M E
Department of Obstetrics & Gynecology, University of Saarland, Saarbrucken, Germany.
FR8.3 Life Science, Department of Genetics & Epigenetics, Saarland University, Saarbrucken, Germany.
Andrologia. 2018 Jan 9. doi: 10.1111/and.12950.
DNA methylation is an epigenetic modification of the genome. The purpose of this study was to determine the influence of cigarette-smoking on sperm DNA methylation from a genomewide survey of sperm samples and to ascertain its effect on sperm parameters. Twenty-eight sperm DNA samples (from 14 fertile smokers as a case study and 14 proven fertile nonsmokers as controls) were subjected to Infinium 450K BeadChip arrays to identify the changes in the DNA methylation level between the two groups. Then, deep bisulphite sequencing was used to validate five CpGs on 78 samples. The results from the Infinium 450K found that only 11 CpGs showed a significant difference in DNA methylation between the case and the control groups. Five CpGs of the eleven (cg00648582, cg0932376, cg19169023, cg23841288 and cg27391564) underwent deep bisulphite sequencing where cg00648582, related to the PGAM5 gene, and the cg23841288 CpGs, related to the PTPRN2 gene amplicons, showed a significant increase in their DNA methylation level in more than one CpG in the case group. In contrast, a significant decrease was found at cg19169023 and at its various neighbouring CpGs in the TYRO3 gene-related amplicons. Furthermore, this study demonstrated a significant correlation between the variation in sperm DNA methylation level and standard sperm parameters in the case group.
DNA甲基化是基因组的一种表观遗传修饰。本研究的目的是通过对精子样本进行全基因组调查，确定吸烟对精子DNA甲基化的影响，并确定其对精子参数的作用。对28个精子DNA样本（14名可育吸烟者作为病例组，14名经证实可育的非吸烟者作为对照组）进行Infinium 450K BeadChip芯片检测，以确定两组之间DNA甲基化水平的变化。然后，使用亚硫酸氢盐深度测序对78个样本中的5个CpG进行验证。Infinium 450K的结果发现，病例组和对照组之间只有11个CpG的DNA甲基化存在显著差异。这11个中的5个CpG（cg00648582、cg0932376、cg19169023、cg23841288和cg27391564）进行了亚硫酸氢盐深度测序，其中与PGAM5基因相关的cg00648582和与PTPRN2基因扩增子相关的cg23841288 CpG在病例组中不止一个CpG位点的DNA甲基化水平显著增加。相反，在与TYRO3基因相关扩增子中的cg19169023及其多个相邻CpG位点发现显著降低。此外，本研究表明病例组中精子DNA甲基化水平的变化与标准精子参数之间存在显著相关性。
