Liu Bang, Cui Yinghong, Chen Wei, Du Li, Li Chunyun, Wan Cailin, He Zuping
The Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, School of Medicine, Hunan Normal University, Changsha, China.
Front Cell Dev Biol. 2021 May 11;9:642916. doi: 10.3389/fcell.2021.642916. eCollection 2021.
Human Sertoli cell is required for completing normal spermatogenesis, and significantly, it has important applications in reproduction and regenerative medicine because of its great plasticity. Nevertheless, the molecular mechanisms underlying the fate decisions of human Sertoli cells remain to be clarified. Here, we have demonstrated the expression, function, and mechanism of Homo sapiens-microRNA (hsa-miR)-100-3p in human Sertoli cells. We revealed that miR-100-3p was expressed at a higher level in human Sertoli cells by 10% fetal bovine serum (FBS) than 0.5% FBS. MiR-100-3p mimics enhanced the DNA synthesis and the proliferation of human Sertoli cells, as indicated by 5-ethynyl-2'-deoxyuridine (EdU) and Cell Counting Kit-8 (CCK-8) assays. Flow cytometry showed that miR-100-3p mimics reduced the apoptosis of human Sertoli cells, and notably, we predicted and further identified serum/glucocorticoid regulated kinase family member 3 (SGK3) as a direct target of MiR-100-3p. SGK3 silencing increased the proliferation and decreased the apoptosis of human Sertoli cells, while SGK3 siRNA 3 assumed a similar role to miR-100-3p mimics in human Sertoli cells. Collectively, our study indicates that miR-100-3p regulates the fate decisions of human Sertoli cells by binding to SGK3. This study is of great significance, since it provides the novel epigenetic regulator for the proliferation and apoptosis of human Sertoli cells and it may offer a new clue for gene therapy of male infertility.
人类支持细胞是完成正常精子发生所必需的，重要的是，由于其具有很大的可塑性，它在生殖和再生医学中具有重要应用。然而，人类支持细胞命运决定的分子机制仍有待阐明。在这里，我们展示了人类微小RNA（hsa-miR）-100-3p在人类支持细胞中的表达、功能和机制。我们发现，与0.5%胎牛血清（FBS）相比，10% FBS培养的人类支持细胞中miR-100-3p表达水平更高。5-乙炔基-2'-脱氧尿苷（EdU）和细胞计数试剂盒-8（CCK-8）检测表明，miR-100-3p模拟物增强了人类支持细胞的DNA合成和增殖。流式细胞术显示，miR-100-3p模拟物减少了人类支持细胞的凋亡，值得注意的是，我们预测并进一步鉴定血清/糖皮质激素调节激酶家族成员3（SGK3）是miR-100-3p的直接靶点。沉默SGK3可增加人类支持细胞的增殖并减少其凋亡，而SGK3 siRNA 3在人类支持细胞中发挥与miR-100-3p模拟物类似的作用。总的来说，我们的研究表明，miR-100-3p通过与SGK3结合来调节人类支持细胞的命运决定。这项研究具有重要意义，因为它为人类支持细胞的增殖和凋亡提供了新的表观遗传调节因子，并且可能为男性不育的基因治疗提供新的线索。
