Center for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China; Center for Reproductive Medicine, Children's Hospital of Shanxi and Women Health Center of Shanxi, Taiyuan, Shanxi, China.
Center for Reproductive Medicine, Children's Hospital of Shanxi and Women Health Center of Shanxi, Taiyuan, Shanxi, China.
Urology. 2022 Jun;164:133-139. doi: 10.1016/j.urology.2022.02.009. Epub 2022 Feb 24.
To explore the candidate genes involved in the pathogenesis of non-obstructive azoospermia (NOA) using bioinformatics analysis and experimental verification.
The gene expression profiles (GSE9210) were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified. We performed function enrichment analyses, constructed protein-protein interaction (PPI) network and identified hub genes. Further, the miRNA-hub genes regulatory network was constructed. Finally, the expression level of CEP55 was verified using RT-qPCR and Western blot, and its diagnostic value was analyzed by the receiver operating characteristic (ROC) curve.
626 DEGs were identified, including 11 upregulated and 615 downregulated genes. Function enrichment analyses showed that these DEGs were significantly enriched in spermatogenesis, fertilization, meiotic cell cycle, flagellated sperm motility, sperm capacitation, spermatid nucleus differentiation and male meiotic nuclear differentiation. The top 10 hub genes were identified including CCNB2, BUB1, TOP2A, BIRC5, CENPF, PBK, NCAPG, DLGAP5, NUF2 and CEP55. In the miRNAs prediction, the hsa-miRNA-449a, hsa-miRNA-34c-5p and hsa-miRNA-34b-5p may be implicated in NOA. In the validation stage, the expression level of CEP55 was significantly decreased in patients with NOA compared to patients with OA. ROC analysis showed that CEP55 had a good diagnostic value for NOA and the combination of CEP55, FSH and mean testicular volume enhanced the prediction performance.
This study identified key genes associated with NOA and their biological functions. Furthermore, CEP55 might play an important role in the pathogenesis of NOA, which will provide novel insights into the targeting therapy of NOA.
利用生物信息学分析和实验验证探讨非梗阻性无精子症（NOA）发病机制中的候选基因。
从基因表达综合数据库（GEO）下载基因表达谱（GSE9210）。鉴定差异表达基因（DEGs）。我们进行了功能富集分析，构建了蛋白质-蛋白质相互作用（PPI）网络，并鉴定了枢纽基因。进一步构建了 miRNA-枢纽基因调控网络。最后，通过 RT-qPCR 和 Western blot 验证了 CEP55 的表达水平，并通过接收者操作特征（ROC）曲线分析了其诊断价值。
鉴定出 626 个 DEGs，包括 11 个上调和 615 个下调基因。功能富集分析表明，这些 DEGs 显著富集于精子发生、受精、减数分裂细胞周期、鞭毛精子运动、精子获能、精子核分化和雄性减数分裂核分化等过程。鉴定出的前 10 个枢纽基因包括 CCNB2、BUB1、TOP2A、BIRC5、CENPF、PBK、NCAPG、DLGAP5、NUF2 和 CEP55。在 miRNA 预测中，hsa-miRNA-449a、hsa-miRNA-34c-5p 和 hsa-miRNA-34b-5p 可能与 NOA 有关。在验证阶段，与 OA 患者相比，NOA 患者的 CEP55 表达水平显著降低。ROC 分析表明，CEP55 对 NOA 具有良好的诊断价值，CEP55、FSH 和平均睾丸体积的联合检测提高了预测性能。
本研究鉴定了与 NOA 相关的关键基因及其生物学功能。此外，CEP55 可能在 NOA 的发病机制中发挥重要作用，为 NOA 的靶向治疗提供新的思路。
