State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Animal Science and Technology, Guangxi University, Nanning, 530004, China.
Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, 528225, China.
Cell Tissue Res. 2023 May;392(2):605-620. doi: 10.1007/s00441-022-03725-7. Epub 2023 Jan 19.
Many studies have shown that circRNAs and miRNAs play important roles in many different life processes. However, the function of circRNAs in spermatogenesis remains unknown. Here, we aimed to explore the mechanisms whereby circRNA-miRNAs-mRNAs regulate abnormal m6A methylation in GC-1spg spermatogonia. We first reduced m6A methylation in GC-1spg whole cells after knocking down the m6A methyltransferase enzyme, METTL3. Then, we performed circRNA- and miRNA-seq on GC-1spg cells with low m6A methylation and identified 48 and 50 differentially expressed circRNAs and miRNAs, respectively. We also predicted the targets of the differentially expressed miRNAs by using Miranda software and further constructed the differentially expressed circRNA-differentially expressed miRNA-mRNA ceRNA network. GO analysis was performed on the differentially expressed circRNAs and miRNA-targeted mRNAs, and an interaction network between the proteins of interest was constructed using Cytoscape. The final GO analysis showed that the target mRNAs were involved in sperm formation. Therefore, a PPI network was subsequently constructed and 2 hub genes (H2afx and Dnmt3a) were identified. In this study, we constructed a ceRNA network and explored the regulatory roles of circRNAs and miRNAs in the pathogenesis of abnormal spermatogenesis caused by low levels of methylated m6A. Also, we identified two pivotal genes that may be key factors in infertility caused by abnormal m6A methylation. This may provide some ideas for the treatment of infertility resulting from abnormal spermatogenesis.
许多研究表明 circRNAs 和 miRNAs 在许多不同的生命过程中发挥着重要作用。然而，circRNAs 在精子发生中的功能仍然未知。在这里，我们旨在探索 circRNA-miRNA-mRNAs 如何调节 GC-1spg 精原细胞中异常的 m6A 甲基化。我们首先在敲低 m6A 甲基转移酶酶 METTL3 后降低 GC-1spg 全细胞中的 m6A 甲基化。然后，我们对 m6A 低甲基化的 GC-1spg 细胞进行了 circRNA 和 miRNA-seq 分析，分别鉴定出 48 个和 50 个差异表达的 circRNAs 和 miRNAs。我们还使用 Miranda 软件预测了差异表达 miRNA 的靶标，并进一步构建了差异表达 circRNA-差异表达 miRNA-mRNA ceRNA 网络。对差异表达 circRNAs 和 miRNA 靶向 mRNAs 进行了 GO 分析，并使用 Cytoscape 构建了感兴趣蛋白的相互作用网络。最终的 GO 分析表明，靶 mRNAs 参与了精子形成。因此，随后构建了一个 PPI 网络，并鉴定出 2 个枢纽基因（H2afx 和 Dnmt3a）。在这项研究中，我们构建了一个 ceRNA 网络，并探讨了 circRNAs 和 miRNAs 在低甲基化 m6A 引起的异常精子发生发病机制中的调节作用。此外，我们鉴定出两个关键基因，它们可能是异常 m6A 甲基化引起的不育的关键因素。这可能为异常精子发生导致的不育症的治疗提供一些思路。
