miR-1306 Mediates the Feedback Regulation of the TGF-/SMAD Signaling Pathway in Granulosa Cells
miR-1306 Mediates the Feedback Regulation of the TGF-/SMAD Signaling Pathway in Granulosa Cells
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miR-1306 介导颗粒细胞中 TGF-/SMAD 信号通路的反馈调节
DOI：
10.3390/cells8040298
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发表时间：
2019-04-01
期刊：
影响因子：
6
通讯作者：
Li, Qifa
中科院分区：
文献类型：
--
作者：
Yang, Liu;Du, Xing;Li, Qifa
转化生长因子-受体II（TGF-β R 2）是TGF-β/SMA-和MAD-相关蛋白（SMAD）信号通路的II型受体，在TGF-β信号转导中起关键作用，并受多种因素调节。然而，参与卵巢中TGFBR 2表达过程的非编码RNA的调节还没有得到很好的研究。在我们的研究中，我们分离并鉴定了猪TGFBR 2基因的3-非翻译区（UTR），并鉴定了microRNA-1306（miR-1306）为靶向猪颗粒细胞（GC）中TGFBR 2的功能性miRNA。功能分析显示，miR-1306促进GC的凋亡以及减弱靶向和损害GC中的TGFBR 2的TGF-/SMAD信号通路。此外，我们鉴定了miR-1306核心启动子，并发现了三个潜在的SMAD 4结合元件（SBE）。荧光素酶和染色质免疫沉淀（ChIP）分析显示，转录因子SMAD 4直接结合miR-1306核心启动子并抑制其转录活性。此外，TGF-/SMAD信号通路通过SMAD 4正反馈经由抑制GC中的miR-1306表达来调节。总的来说，我们的研究结果提供了证据的表观遗传机制，调制以及介导的反馈调节的经典TGF-/SMAD信号通路在GC从猪卵巢。
Transforming growth factor- receptor II (TGFBR2), the type II receptor of the TGF-/SMA- and MAD-related protein (SMAD) signaling pathway, plays a crucial role in TGF- signal transduction and is regulated by multiple factors. Nevertheless, the modulation of the non-coding RNA involved in the process of TGFBR2 expression in ovaries is not well studied. In our study, we isolated and characterized the 3-untranslated region (UTR) of the porcine TGFBR2 gene and microRNA-1306 (miR-1306) was identified as the functional miRNA that targets TGFBR2 in porcine granulosa cells (GCs). Functional analysis showed that miR-1306 promotes apoptosis of GCs as well as attenuating the TGF-/SMAD signaling pathway targeting and impairing TGFBR2 in GCs. Moreover, we identified the miR-1306 core promoter and found three potential SMAD4-binding elements (SBEs). Luciferase and chromatin immunoprecipitation (ChIP) assays revealed that the transcription factor SMAD4 directly binds to the miR-1306 core promoter and inhibits its transcriptional activity. Furthermore, the TGF-/SMAD signaling pathway is modulated by SMAD4 positive feedback via inhibition of miR-1306 expression in GCs. Collectively, our findings provide evidence of an epigenetic mechanism that modulates as well as mediates the feedback regulation of the classical TGF-/SMAD signaling pathway in GCs from porcine ovaries.