Momeni Abdolmabood, Najafipour Reza, Hamta Ahmad, Jahani Suzan, Moghbelinejad Sahar
Cellular and Molecular Research Centre, Qazvin University of Medical Sciences, P.O. Box: 341197-5981, Qazvin, Iran.
Biology Department, School of Basic Science Arak University, Arak, Iran.
Reprod Sci. 2020 Jan;27(1):301-308. doi: 10.1007/s43032-019-00025-4. Epub 2020 Jan 1.
Production of high-quality spermatozoa is necessary for male fertility. In this regard, post-mitotic stage in spermatogenesis is very important which posttranscriptional microRNAs (miRNAs) playing a key role at this stage. In this research, we evaluated the expression and methylation of hsa-miR-34 family in sperm samples of infertile men. We recruited 102 infertile men (asthenozoospermia, teratozoospermia, asthenoteratozoospermia, and oligoasthenoteratozoospermia) and 52 fertile men as control. The expression of hsa-miR-34a,b,c was determined by quantitative real-time PCR (qRT-PCR) technique. Methylation of hsa-miR-34b,c promoter was evaluated by methylation-specific PCR (MS-PCR) method. Our data indicated under-expression of three miRNAs (hsa-miR-34a,b,c) in the sperm samples of infertile men in compared to their fertile counterparts. The highest rate of expression reduction was observed in hsa-miR-34c-5p and in oligoasthenoteratospermic patients (P = 0.011, F = 4.01). The results revealed that the frequency of methylation of the promoter region of hsa-miR-34b,c in infertile men was higher than that of fertile men (82.4% versus 23.3%), and the highest frequency of methylation was observed in patients with asthenoteratospermia (92.9%) and oligoasthenoteratospermia (93.8%). In conclusion, our results indicated lower expression of hsa-miR-34a,b,c and hypermethylation of hsa-miR-34b,c promoter in sperm samples of infertile men. Aberrant under-expression of these miRNAs could be duo to the hypermethylation of the promoter region and indicative of a defect in spermatogenesis.
产生高质量的精子是男性生育能力所必需的。在这方面，精子发生的有丝分裂后阶段非常重要，转录后微小RNA（miRNA）在该阶段发挥关键作用。在本研究中，我们评估了不育男性精子样本中hsa-miR-34家族的表达和甲基化情况。我们招募了102名不育男性（弱精子症、畸形精子症、弱畸精子症和少弱畸精子症）和52名生育能力正常的男性作为对照。通过定量实时PCR（qRT-PCR）技术测定hsa-miR-34a、b、c的表达。通过甲基化特异性PCR（MS-PCR）方法评估hsa-miR-34b、c启动子的甲基化情况。我们的数据表明，与生育能力正常的男性相比，不育男性精子样本中三种miRNA（hsa-miR-34a、b、c）表达不足。在hsa-miR-34c-5p和少弱畸精子症患者中观察到最高的表达降低率（P = 0.011，F = 4.01）。结果显示，不育男性中hsa-miR-34b、c启动子区域的甲基化频率高于生育能力正常的男性（82.4%对23.3%），在弱畸精子症患者（92.9%）和少弱畸精子症患者（93.8%）中观察到最高的甲基化频率。总之，我们的结果表明不育男性精子样本中hsa-miR-34a、b、c表达较低，hsa-miR-34b、c启动子存在高甲基化。这些miRNA的异常低表达可能是由于启动子区域的高甲基化，表明精子发生存在缺陷。
