Immature Sertoli cell proliferation determines the total number of mature Sertoli cells and further regulates normal spermatogenesis. Accumulating evidence demonstrates that microRNAs (miRNAs) play regulatory roles in immature Sertoli cell proliferation, while the functions and mechanisms of the Sertoli cells of domestic animals are poorly understood. In the present study, we aimed to investigate the roles of miR-362 in cell proliferation and apoptosis of porcine immature Sertoli cells. The results showed that miR-362 inhibition promoted the entrance of cells into the S phase and increased the expressions of cell cycle-related genes c-MYC, CNNE1, CCND1 and CDK4. Knock-down of miR-362 also promoted cell proliferation and inhibited apoptosis, which was demonstrated by the results from cell counting kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) and Annexin V-FITC/PI staining assays. The recQ-mediated genome instability protein 1 (RMI1) gene was identified as a potential target gene of miR-362 via luciferase reporter assay, and miR-362 repressed the protein expression of RMI1 in porcine immature Sertoli cells. siRNA-induced RMI1 knock-down further abolished the effects of miR-362 inhibition on porcine immature Sertoli cells. Collectively, we concluded that miR-362 knock-down promotes proliferation and inhibits apoptosis in porcine immature Sertoli cells by targeting the RMI1 gene, which indicates that miR-362 determines the fate of immature Sertoli cells.

中文翻译：

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通过靶向RMI1基因，miR-362敲低可促进猪未成熟Sertoli细胞的增殖并抑制其凋亡。

Sertoli细胞的未成熟增殖决定了成熟Sertoli细胞的总数，并进一步调节正常的精子发生。越来越多的证据表明，微小RNA（miRNA）在未成熟的Sertoli细胞增殖中起调节作用，而对家畜的Sertoli细胞的功能和机制却知之甚少。在本研究中，我们旨在研究miR-362在猪未成熟Sertoli细胞增殖和凋亡中的作用。结果表明，miR-362抑制促进细胞进入S期并增加细胞周期相关基因c-MYC，CNNE1，CCND1和CDK4的表达。miR-362的抑制还促进细胞增殖并抑制细胞凋亡，这由细胞计数试剂盒8（CCK-8），5-乙炔基2'的结果证明。-脱氧尿苷（EdU）和膜联蛋白V-FITC / PI染色测定。通过荧光素酶报告基因测定，recQ介导的基因组不稳定性蛋白1（RMI1）基因被鉴定为miR-362的潜在靶基因，而miR-362抑制了RMI1在猪未成熟Sertoli细胞中的蛋白表达。siRNA诱导的RMI1敲低进一步消除了miR-362抑制对猪未成熟Sertoli细胞的影响。总的来说，我们得出的结论是，miR-362敲低可以通过靶向RMI1基因来促进猪未成熟Sertoli细胞的增殖并抑制其凋亡，这表明miR-362决定了未成熟Sertoli细胞的命运。miR-362可抑制猪未成熟Sertoli细胞中RMI1的蛋白表达。siRNA诱导的RMI1敲低进一步消除了miR-362抑制对猪未成熟Sertoli细胞的影响。总的来说，我们得出的结论是，miR-362敲低可以通过靶向RMI1基因来促进猪未成熟Sertoli细胞的增殖并抑制其凋亡，这表明miR-362决定了未成熟Sertoli细胞的命运。miR-362可抑制猪未成熟Sertoli细胞中RMI1的蛋白表达。siRNA诱导的RMI1敲低进一步消除了miR-362抑制对猪未成熟Sertoli细胞的影响。总的来说，我们得出的结论是，miR-362敲低可以通过靶向RMI1基因来促进猪未成熟Sertoli细胞的增殖并抑制其凋亡，这表明miR-362决定了未成熟Sertoli细胞的命运。