Air pollution and the application of Silica nanoparticles (SiNPs) have increased the risk of human exposure to SiNPs. SiNPs are known to induce cytotoxicity in spermatocyte cells (GC-2spd cells) of mice and male reproductive system damage. However, the expression profiles of miRNA and mRNA and the molecular mechanism of miRNA-mRNA integration in reproductive toxicity induced by SiNPs in GC-2spd cells are still unclear. Therefore, GC-2spd cells were divided into 0 μg/mL and 5 μg/mL SiNPs groups, and the cells were collected and analyzed after passaging for 30 generations using miRNA microarray and Illumina high-throughput sequencing (Illumina HiSeq) for the integrated analysis of miRNA and mRNA expression. Both miRNA Microarray and Illumina Hiseq identified 15 significant differentially expressed miRNAs and 1648 significant differentially expressed mRNAs. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and miRNA-gene-pathway-network analysis revealed 15 significant differentially expressed miRNAs that could regulate the DNA replication and the fatty acid metabolism, respectively. Furthermore, the mRNA-mRNA regulatory network analysis revealed that Pkfl (phosphofructokinase, liver, B-type) and DHCR24 (24-dehydrocholesterol reductase) were highly expressed, but also affected DNA replication and fatty acid metabolism in SiNPs-treated GC-2spd cells. Additionally, miRNA-mRNA integration analysis revealed that miRNA-138-1-3p might have a regulatory relationship with fatty acid metabolism and DNA replication. It is confirmed that SiNPs could decrease the expression of 10 miRNAs and increase the expression of 5 miRNAs. These findings suggest that the cytotoxicity of GC-2spd cells induced by SiNPs depends on the deregulation of multiple miRNAs, which regulate the DNA replication and fatty acid metabolism. Our results are the first to establish an integrated analysis of miRNA–mRNA interactions and mRNA-mRNA and defines multiple pathways involved in SiNPs-treated GC-2spd cells.

空气污染和二氧化硅纳米粒子 (SiNPs) 的应用增加了人类暴露于 SiNPs 的风险。已知 SiNP 在小鼠的精母细胞（GC-2spd 细胞）中诱导细胞毒性和雄性生殖系统损伤。然而，miRNA和mRNA的表达谱以及miRNA-mRNA整合在SiNPs诱导GC-2spd细胞生殖毒性中的分子机制仍不清楚。因此，将GC-2spd细胞分为0 μg/mL和5 μg/mL SiNPs组，传代30代后收集细胞并使用miRNA芯片和Illumina高通量测序（Illumina HiSeq）进行整合分析miRNA 和 mRNA 的表达。miRNA 微阵列和 Illumina Hiseq 均鉴定了 15 个显着差异表达的 miRNA 和 1648 个显着差异表达的 mRNA。基因本体论 (GO) 富集分析、京都基因和基因组百科全书 (KEGG) 通路富集分析和 miRNA-基因-通路-网络分析揭示了 15 个显着差异表达的 miRNA，它们分别可以调节 DNA 复制和脂肪酸代谢。此外，mRNA-mRNA 调控网络分析显示，Pkfl（磷酸果糖激酶，肝脏，B 型）和 DHCR24（24-脱氢胆固醇还原酶）高表达，但也影响 SiNPs 处理的 GC-2spd 细胞中的 DNA 复制和脂肪酸代谢. 此外，miRNA-mRNA整合分析表明，miRNA-138-1-3p可能与脂肪酸代谢和DNA复制有调节关系。已证实SiNPs可以降低10个miRNA的表达并增加5个miRNA的表达。这些发现表明，SiNPs 诱导的 GC-2spd 细胞的细胞毒性取决于多种 miRNAs 的失调，这些 miRNAs 调节 DNA 复制和脂肪酸代谢。我们的结果首次建立了对 miRNA-mRNA 相互作用和 mRNA-mRNA 的综合分析，并定义了 SiNPs 处理的 GC-2spd 细胞中涉及的多种途径。这些发现表明，SiNPs 诱导的 GC-2spd 细胞的细胞毒性取决于多种 miRNAs 的失调，这些 miRNAs 调节 DNA 复制和脂肪酸代谢。我们的结果首次建立了对 miRNA-mRNA 相互作用和 mRNA-mRNA 的综合分析，并定义了 SiNPs 处理的 GC-2spd 细胞中涉及的多种途径。这些发现表明，SiNPs 诱导的 GC-2spd 细胞的细胞毒性取决于多种 miRNAs 的失调，这些 miRNAs 调节 DNA 复制和脂肪酸代谢。我们的结果首次建立了对 miRNA-mRNA 相互作用和 mRNA-mRNA 的综合分析，并定义了 SiNPs 处理的 GC-2spd 细胞中涉及的多种途径。