MicroRNA-449a Suppresses Mouse Spermatogonia Proliferation via Inhibition of CEP55
MicroRNA-449a Suppresses Mouse Spermatogonia Proliferation via Inhibition of CEP55
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MicroRNA-449a 通过抑制 CEP55 抑制小鼠精原细胞增殖
DOI：
10.1007/s43032-020-00354-9
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发表时间：
2020-10-23
影响因子：
2.9
通讯作者：
Zhu, Yong-tong
中科院分区：
文献类型：
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作者：
Hua, Rui;Chu, Qing-jun;Zhu, Yong-tong
目前，患有成熟停滞（MA）的不育患者很难获得成熟精子。精子发生及其分子机制尚不清楚。收集MA和正常精子发生（NS）的患者。采用iTRAQ蛋白质组学方法分析了它们之间的差异蛋白质。为了验证蛋白质组数据的置信度，通过蛋白质印迹法（WB）、定量聚合酶链反应（qPCR）和免疫荧光法分析单个样品。通过荧光素酶测定法测定miR-449 a和CEP 55。用CEP 55 siRNA、miR-449 a模拟物或抑制剂转染小鼠GC-1细胞，并测定细胞增殖。与正常对照组相比，MA组有27个蛋白质表达差异，其中CEP 55蛋白差异最显著。WB和qPCR结果显示，与MA组相比，NS组CEP 55表达水平明显升高。在转染的细胞中，miR-449 a的过表达和CEP 55的敲低均下调CEP 55的表达并降低细胞增殖。miR-449 a通过抑制CEP 55抑制小鼠精原细胞增殖。
At present, infertile patients with maturation arrest (MA) are difficult to obtain mature sperm. Spermatogenesis and its molecular mechanism are still not clear. Patients with MA and normal spermatogenesis (NS) were collected. iTRAQ-based proteomic approach was performed to reveal the different proteins between them. To validate the confidence of proteome data, the individual samples were analyzed by Western blotting (WB), quantitative polymerase chain reaction (qPCR), and immunofluorescence. The miR-449a and CEP55 were determined by Luciferase assay. Mouse GC-1 cells were transfected with CEP55 siRNAs, miR-449a mimic, or inhibitor, and cell proliferation was determined. Compared with NS, 27 proteins were differentially expressed in MA, and CEP55 protein was the most significant difference. WB and qPCR showed that CEP55 levels were significantly elevated in NS than MA. In transfected cells, overexpression of miR-449a and knockdown of CEP55 both downregulated CEP55 expression and decreased cell proliferation. miR-449a suppresses mouse spermatogonia proliferation via inhibition of CEP55.