miR-106a Increases Granulosa Cell Viability and Is Downregulated in Women With Diminished Ovarian Reserve
miR-106a Increases Granulosa Cell Viability and Is Downregulated in Women With Diminished Ovarian Reserve
复制标题
miR-106a 增加颗粒细胞活力，并在卵巢储备减少的女性中下调
DOI：
10.1210/jc.2017-02344
复制
发表时间：
2018-06-01
影响因子：
5.8
通讯作者：
Yang, Xiaokui
中科院分区：
文献类型：
--
作者：
Hong, Liming;Peng, Sha;Yang, Xiaokui
上下文
卵巢储备功能下降的妇女生育能力降低，心血管事件和骨质疏松症。尽管在几种卵巢疾病中已经描述了差异microRNA（miRNA）表达，但关于miRNA在DOR发病机制中的作用知之甚少。
目的
鉴定DOR中差异表达的miRNAs并探讨miR-106a在人类颗粒细胞增殖中的作用。
设计
采用miRNA微阵列（n = 3）和定量逆转录聚合酶链反应（n = 30）检测正常周期和DOR妇女血清和颗粒细胞中miRNA的表达。在评估细胞活力和凋亡之前，单独或与miR-106 a模拟物、miR-106 a抑制剂、凋亡信号调节激酶1（ASK 1）小干扰RNA（siRNA）或p38丝裂原活化蛋白激酶（MAPK）抑制剂（SB 203580）组合处理原代人颗粒细胞。Western blot检测ASK1蛋白表达及p38 MAPK磷酸化/活化。通过3 '非翻译区（3'UTR）荧光素酶分析检查miR-106a与ASK 1 mRNA的结合。
结果
15个miRNAs差异表达（n = 30），miR-106a在DOR妇女血清和颗粒细胞中下调。miR-106a模拟物增加了细胞活力并减弱了凋亡，而用miR-106a抑制剂处理后发生了相反的情况。miR-106a通过直接靶向其3'UTR抑制ASK 1表达。miR-106a抑制剂增加p38 MAPK磷酸化/活化，并且这种作用被ASK1 siRNA处理消除。尽管ASK1的敲低消除了miR-106a抑制剂对细胞活力/凋亡的影响，但用SB 203580预处理并没有显著改变miR-106a抑制剂的作用。
结论
miR-106a的下调可能通过降低颗粒细胞活力和通过增强的ASK 1信号促进细胞凋亡而促进DOR的发病机制。
Context
Women with diminished ovarian reserve (DOR) have reduced fertility, cardiovascular events, and osteoporosis. Although differential microRNA (miRNA) expression has been described in several ovarian disorders, little is known about the role of miRNAs in the pathogenesis of DOR.
Objective
Identify differentially expressed miRNAs in DOR and explore the role of miR-106a in human granulosa cell proliferation.
Design
miRNA microarray (n = 3) and quantitative reverse transcription polymerase chain reaction (n = 30) were used to examine miRNA expression in serum and granulosa cells from normal-cycling and women with DOR. Primary human granulosa cells were treated alone or in combination with miR-106a mimic, miR-106a inhibitor, apoptosis signal-regulating kinase 1 (ASK1) small interfering RNA (siRNA), or p38 mitogen-activated protein kinase (MAPK) inhibitor (SB203580) before assessment of cell viability and apoptosis. Western blot was used to measure ASK1 protein and phosphorylation/activation of p38 MAPK. Binding of miR-106a to ASK1 mRNA was examined by 3' untranslated region (3'UTR) luciferase analysis.
Results
Fifteen miRNAs were differentially expressed (n = 30), and miR-106a was downregulated in serum and granulosa cells of women with DOR. miR-106a mimic increased cell viability and attenuated apoptosis, whereas the converse occurred following treatment with miR-106a inhibitor. miR-106a suppressed ASK1 expression by directly targeting its 3'UTR. miR-106a inhibitor increased p38 MAPK phosphorylation/activation, and this effect was abolished by treatment with ASK1 siRNA. Whereas knockdown of ASK1 abolished the effects of miR-106a inhibitor on cell viability/apoptosis, pretreatment with SB203580 did not significantly alter the effects of miR-106a inhibitor.
Conclusions
Downregulation of miR-106a may contribute to the pathogenesis of DOR by reducing granulosa cell viability and promoting apoptosis via enhanced ASK1 signaling.