Li Zhiming, Zhuang Xuan, Zeng Jinxiong, Tzeng Chi-Meng
Translational Medicine Research Center, School of Pharmaceutical Sciences, Xiamen UniversityXiamen China.
Department of Pathology, Wake Forest University School of MedicineWinston-Salem, NC, USA.
Front Physiol. 2017 May 12;8:261. doi: 10.3389/fphys.2017.00261. eCollection 2017.
Severe oligozoospermia (SO) is a complex disorder, whose etiology is the combined effect of genetic factors and epigenetic conditions. In this study, we examined DNA methylation and mRNA expression status in a set of testicular tissues of SO patients ( = 3), and compared methylated data with those derived from obstructive azoospermia (OA) patients ( = 3) with normal spermatogenesis phenotype. We identified 1,960 differentially methylated CpG sites showing significant alterations in SO vs. OA using the Illumina Infinium HumanMethylation450 bead array. By integrating above DNA methylation data and mRNA expression results, we totally identified 72 methylated CpG sites located in 65 genes with anti-correlation between DNA methylation and mRNA expression. Integrated pathways analysis indicates that these genes are involved in response to hormone stimulus, activation of protein kinase activity, and apoptotic process, among others. We also observed some genes with inversely correlated difference is novel in male infertility field, including PTPRN2, EPHX1, SERPINB9, SLIT3, etc. Our results lay a groundwork for further biological study of SO. Moreover, we generated a workflow for integrated analysis of DNA methylation and mRNA expression, which is expandable to other study types.
严重少精子症（SO）是一种复杂的疾病，其病因是遗传因素和表观遗传条件共同作用的结果。在本研究中，我们检测了一组SO患者（n = 3）睾丸组织中的DNA甲基化和mRNA表达状态，并将甲基化数据与具有正常精子发生表型的梗阻性无精子症（OA）患者（n = 3）的数据进行了比较。我们使用Illumina Infinium HumanMethylation450 Bead芯片鉴定出1960个差异甲基化的CpG位点，这些位点在SO与OA中显示出显著变化。通过整合上述DNA甲基化数据和mRNA表达结果，我们总共鉴定出72个位于65个基因中的甲基化CpG位点，其DNA甲基化与mRNA表达呈负相关。综合通路分析表明，这些基因参与激素刺激反应、蛋白激酶活性激活和凋亡过程等。我们还观察到一些在男性不育领域中差异呈负相关的新基因，包括PTPRN2、EPHX1、SERPINB9、SLIT3等。我们的结果为SO的进一步生物学研究奠定了基础。此外，我们生成了一个用于DNA甲基化和mRNA表达综合分析的工作流程，该流程可扩展到其他研究类型。
