LncRNA-Gm2044 is transcriptionally activated by A-MYB and regulates Sycp1 expression as a miR-335-3p sponge in mouse spermatocyte-derived GC-2spd(ts) cells
LncRNA-Gm2044 is transcriptionally activated by A-MYB and regulates Sycp1 expression as a miR-335-3p sponge in mouse spermatocyte-derived GC-2spd(ts) cells
复制标题
LncRNA-Gm2044 由 A-MYB 转录激活，并在小鼠精母细胞衍生的 GC-2spd(ts) 细胞中作为 miR-335-3p 海绵调节 Sycp1 表达
DOI：
10.1016/j.diff.2020.05.004
复制
发表时间：
2020-07-01
期刊：
影响因子：
2.9
通讯作者：
Hu, Ke
中科院分区：
文献类型：
--
作者：
Liang, Meng;Wang, Haiyan;Hu, Ke
长链非编码RNA（lncRNA）在精子发生中起着重要作用。然而，很少有人知道如何lncRNA基因的表达本身在睾丸生殖细胞的调节。我们先前证明lncRNA-Gm 2044在精母细胞中存在高表达，并且可以调节雄性生殖细胞的增殖。本研究进一步探讨了lnRNA-Gm 2044在精母细胞中表达的转录调控及其下游信号传导机制。生物信息学评估预测了lncRNA-Gm 2044启动子区域中精母细胞特异性转录因子A-MYB的两个潜在结合位点。结果表明，转录因子A-MYB可促进小鼠精母细胞来源的GC-2spd（ts）细胞中lncRNA-Gm 2044的表达。ChIP和荧光素酶检测证实，A-MYB主要与lncRNA-Gm 2044的启动子远端（距转录起始位点约819 bp）结合，并通过该位点调控lncRNA-Gm 2044的表达。此外，我们证实lncRNA-Gm 2044作为miR-335- 3 p海绵发挥作用，以提高miR-335- 3 p的直接靶蛋白Sycp 1的水平。此外，A-MYB可通过激活其靶点lncRNA-Gm 2044上调Sycp 1表达并下调GC-2spd（ts）细胞增殖。lncRNA-Gm 2044的过表达或miR-335- 3 p的敲减可以部分地挽救A-MYB对Sycp 1表达和GC-2spd（ts）细胞增殖的影响，这些结果为lncRNA-miRNA在转录因子A-MYB调控精母细胞功能中的作用机制提供了新的信息。
Long noncoding RNAs (lncRNAs) have been shown to execute key roles in spermatogenesis. However, little is known about how lncRNAs gene expression is itself regulated in the germ cells of testis. We previously demonstrated that high expression of lncRNA-Gm2044 exists in spermatocytes and can regulate male germ cell proliferation. Here, the transcriptional regulation of lnRNA-Gm2044 expression in spermatocytes and the downstream signaling were further explored. A bioinformatics assessment predicted two potential binding-sites for the spermatocyte-specific transcription factor A-MYB in the promoter region of lncRNA-Gm2044. Our results proved that the transcription factor A-MYB promotes the expression of lncRNA-Gm2044 in mouse spermatocyte-derived GC-2spd(ts) cells. ChIP and luciferase assays verified that A-MYB mainly binds to the distal promoter region (-819 bp relative to the transcription start site) of lncRNA-Gm2044 and regulates lncRNA-Gm2044 expression through the -819 bp binding-site. In addition, we confirmed that lncRNA-Gm2044 functions as a miR-335-3p sponge to enhance the levels of miR-335-3p's direct target protein, Sycp1. Furthermore, A-MYB can upregulate Sycp1 expression and down-regulate GC-2spd(ts) cell proliferation by activating its target, lncRNA-Gm2044. Overexpression of lncRNA-Gm2044 or knockdown of miR-335-3p can, at least partially, rescue the effects of A-MYB on Sycp1 expression and GC-2spd(ts) cell proliferation.Taken together, our results provide new information on the mechanistic roles of lncRNA-miRNA in transcription factor A-MYB regulation of spermatocyte function.