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不育男性精子样本中hsa-miR-34家族的表达及甲基化模式

Expression and Methylation Pattern of hsa-miR-34 Family in Sperm Samples of Infertile Men.

作者信息

Momeni Abdolmabood, Najafipour Reza, Hamta Ahmad, Jahani Suzan, Moghbelinejad Sahar

机构信息

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.

出版信息

Reprod Sci. 2020 Jan;27(1):301-308. doi: 10.1007/s43032-019-00025-4. Epub 2020 Jan 1.

Abstract

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.

摘要

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。

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