Li Zhiming, Zheng Zaozao, Ruan Jun, Li Zhi, Zhuang Xuan, Tzeng Chi-Meng
Translational Medicine Research Center (TMRC), School of Pharmaceutical Sciences, Xiamen University, Xiamen, Fujian, China.
Key Laboratory for Cancer T-Cell Theranostics and Clinical Translation (CTCTCT), Xiamen University, Xiamen, Fujian, China.
Oncotarget. 2016 Aug 16;7(33):52781-52796. doi: 10.18632/oncotarget.10947.
Our previous research suggested that an integrated analysis of microRNA (miRNA) and messenger RNA (mRNA) expression is helpful to explore miRNA-mRNA interactions and to uncover the molecular mechanisms of male infertility. In this study, microarrays were used to compare the differences in the miRNA and mRNA expression profiles in the testicular tissues of severe oligozoospermia (SO) patients with obstructive azoospermia (OA) controls with normal spermatogenesis. Four miRNAs (miR-1246, miR-375, miR-410, and miR-758) and six mRNAs (SLC1A3, PRKAR2B, HYDIN, WDR65, PRDX1, and ADATMS5) were selected to validate the microarray data using quantitative real-time PCR. Using statistical calculations and bioinformatics predictions, we identified 33 differentially expressed miRNAs and 1,239 differentially expressed mRNAs, among which one potential miRNA-target gene pair, miR-34c-3p and PLCXD3 (Phosphatidylinositol-Specific Phospholipase C, X Domain Containing 3), was identified. Immunohistochemical analysis indicated that PLCXD3 was located within the germ cells of the mouse and human testis. Moreover, we found that miR-34c-3p was able to decrease PLCXD3 expression in mouse (GC-1 and TM4) and human (NCM460) cell lines, presumably indicating the possibility that miR-34c-3p acts as an intracellular mediator in germinal lineage differentiation. Notably, we reported the expression of the PLCXD3 protein in a man with normal spermatogenesis and the lack of the PLCXD3 protein in a man with SO. Therefore, the identified miRNA and mRNA may represent a potentially novel molecular regulatory network and therapeutic targets for the study or treatment of SO, which might provide a better understanding of the molecular basis of spermatogenesis dysfunction.
我们之前的研究表明，对微小RNA（miRNA）和信使RNA（mRNA）表达进行综合分析，有助于探索miRNA-mRNA相互作用，并揭示男性不育的分子机制。在本研究中，我们使用微阵列比较了严重少精子症（SO）患者与梗阻性无精子症（OA）对照（精子发生正常）睾丸组织中miRNA和mRNA表达谱的差异。选择了4种miRNA（miR-1246、miR-375、miR-410和miR-758）和6种mRNA（SLC1A3、PRKAR2B、HYDIN、WDR65、PRDX1和ADATMS5），通过定量实时PCR验证微阵列数据。通过统计计算和生物信息学预测，我们鉴定出33种差异表达的miRNA和1239种差异表达的mRNA，其中鉴定出一个潜在的miRNA-靶基因对，即miR-34c-3p和PLCXD3（含X结构域的磷脂酰肌醇特异性磷脂酶C 3）。免疫组织化学分析表明，PLCXD3定位于小鼠和人类睾丸的生殖细胞内。此外，我们发现miR-34c-3p能够降低小鼠（GC-1和TM4）和人类（NCM460）细胞系中PLCXD3的表达，这可能表明miR-34c-3p作为生殖细胞系分化的细胞内介质的可能性。值得注意的是，我们报告了PLCXD3蛋白在精子发生正常的男性中的表达情况，以及在少精子症男性中PLCXD3蛋白的缺失情况。因此，鉴定出的miRNA和mRNA可能代表了一个潜在的新型分子调控网络以及少精子症研究或治疗的治疗靶点，这可能有助于更好地理解精子发生功能障碍的分子基础。
