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类固醇生成因子 1（SF1）对 miR-202-5p 的转录激活通过靶向 TGFβR2 诱导山羊颗粒细胞凋亡。

Transactivation of miR-202-5p by Steroidogenic Factor 1 (SF1) Induces Apoptosis in Goat Granulosa Cells by Targeting TGFβR2.

机构信息

Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.

出版信息

Cells. 2020 Feb 14;9(2):445. doi: 10.3390/cells9020445.

DOI:10.3390/cells9020445
PMID:32075111
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7072820/
Abstract

MicroRNAs play key roles during ovary development, with emerging evidence suggesting that miR-202-5p is specifically expressed in female animal gonads. Granulosa cells (GCs) are somatic cells that are closely related to the development of female gametes in mammalian ovaries. However, the biological roles of miR-202-5p in GCs remain unknown. Here, we show that miR-202-5p is specifically expressed in GCs and accumulates in extracellular vesicles (EVs) from large growth follicles in goat ovaries. In vitro assays showed that miR-202-5p induced apoptosis and suppressed the proliferation of goat GCs. We further revealed that miR-202-5p is a functional miRNA that targets the transforming growth factor-beta type II receptor (). MiR-202-5p attenuated TGF-β/SMAD signaling through the degradation of TGFβR2 at both the mRNA and protein level, decreasing p-SMAD3 levels in GCs. Moreover, we verified that steroidogenic factor 1 (SF1) is a transcriptional factor that binds to the promoters of miR-202 and cytochrome P450 family 19 subfamily A member 1 () through luciferase reporter and chromatin immunoprecipitation (ChIP) assays. That contributed to positive correlation between miR-202-5p and expression and estradiol (E2) release. Furthermore, SF1 repressed TGFβR2 and p-SMAD3 levels in GCs through the transactivation of miR-202-5p. Taken together, these results suggest a mechanism by which miR-202-5p regulates canonical TGF-β/SMAD signaling through targeting in GCs. This provides insight into the transcriptional regulation of and during goat ovarian follicular development.

摘要

微小 RNA 在卵巢发育过程中发挥关键作用，有新证据表明 miR-202-5p 特异性表达于雌性动物性腺。颗粒细胞 (GCs) 是与哺乳动物卵巢中雌性配子发育密切相关的体细胞。然而，miR-202-5p 在 GCs 中的生物学作用尚不清楚。在这里，我们表明 miR-202-5p 特异性表达于 GCs，并在山羊卵巢大生长卵泡的细胞外囊泡 (EVs) 中积累。体外实验表明，miR-202-5p 诱导 GC 凋亡并抑制其增殖。我们进一步揭示了 miR-202-5p 是一种功能性 miRNA，可靶向转化生长因子-β 型 II 受体 (TGFβR2)。miR-202-5p 通过降解 TGFβR2 的 mRNA 和蛋白水平，减弱 TGF-β/SMAD 信号通路，降低 GC 中的 p-SMAD3 水平。此外，我们验证了类固醇生成因子 1 (SF1) 通过荧光素酶报告和染色质免疫沉淀 (ChIP) 实验是结合到 miR-202 和细胞色素 P450 家族 19 亚家族 A 成员 1 () 启动子上的转录因子。这有助于 miR-202-5p 与表达和雌二醇 (E2) 释放之间的正相关。此外，SF1 通过 miR-202-5p 的反式激活抑制 GC 中的 TGFβR2 和 p-SMAD3 水平。总之，这些结果表明，miR-202-5p 通过靶向 GCs 中的 TGFβR2 调节经典 TGF-β/SMAD 信号通路的机制。这为山羊卵巢卵泡发育过程中对和的转录调控提供了新的认识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/6900d76e86e4/cells-09-00445-g007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/3e2c205c6090/cells-09-00445-g005.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/6900d76e86e4/cells-09-00445-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/5c2aefcf3b70/cells-09-00445-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/a4f17e676eeb/cells-09-00445-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/3eb03f842e38/cells-09-00445-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/0b6d56eeba8f/cells-09-00445-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/3e2c205c6090/cells-09-00445-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/eecfeae2d4a9/cells-09-00445-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e42/7072820/6900d76e86e4/cells-09-00445-g007.jpg

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