miR-196a overexpression activates the MEK/ERK signaland represses the progesterone receptor and decidualizationin eutopic endometrium from women with endometriosis.
miR-196a overexpression activates the MEK/ERK signaland represses the progesterone receptor and decidualizationin eutopic endometrium from women with endometriosis.
复制标题
miR-196a 过表达激活 MEK/ERK 信号并抑制子宫内膜异位症女性在位子宫内膜中的孕酮受体和蜕膜化。
DOI：
10.1093/humrep/dew223
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发表时间：
2016
影响因子：
6.1
通讯作者：
Huang Wei
中科院分区：
文献类型：
--
作者：
Zhou Min;Fu Jing;Xiao Li;Yang Shiyuan;Song Yong;Zhang Xianghui;Feng Xue;Sun Huaqin;Xu Wenming;Huang Wei
研究问题microRNAs（miRNAs）是否参与子宫内膜异位症患者在位内膜中孕激素受体（PGR）的异常表达？miR-196 a上调MEK/ERK信号传导，介导轻度或轻微子宫内膜异位症患者在位子宫内膜中PGR表达下调。已知植入失败被强烈认为是轻度或轻微子宫内膜异位症患者不孕的潜在原因。孕激素抵抗主要是由在位子宫内膜中孕激素受体的异常表达引起的，被认为是子宫内膜容受性降低的关键因素;迄今为止，表观遗传学而不是miRNA已被证明影响PGR在子宫内膜中的表达。验证了miR-196 a的差异表达。生物信息学分析预测miR-196 a靶向PCR的3′-非翻译区（UTR）。miR-196 a水平和PGR表达之间的关系进行了研究和MEK/ERK信号通路的作用investigated.Participants/MATERIALS，设置，METHODS总RNA提取自在位子宫内膜样本中的三个不孕妇女轻度/最小子宫内膜异位症和三个无疾病的对照科目。通过微阵列分析来分析miRNA和mRNA表达水平。通过qRT-PCR验证miR-196 a表达[子宫内膜异位症（n= 22）和对照（n= 20）]，而功能分析利用体外转染子宫内膜基质细胞（ESC），诱导ESC的蜕膜化，主要结果和CHANCEA的作用通过微阵列筛选总共66个失调的miRNA和357个失调的mRNA分析. miR-196 a和P-MEK/P-ERK在轻度/极轻度子宫内膜异位症患者的在位子宫内膜中均显著上调。PGR和PGR-B mRNA被miR-196 a过表达抑制，并被miR-196 a抑制上调。荧光素酶报告基因未能证实miR-196 a对PCR的靶向调控。用miR-196 a模拟物转染ESC导致P-MEK/P-ERK蛋白水平增加，PGR蛋白水平降低和非典型蜕膜化。在miR-196 a抑制后，P-MEK/P-ERK蛋白下调，PGR蛋白上调。抑制P-MEK/P-ERK也能增加PGR的表达。大规模数据数据见补充表SI和SII。限制性因素抑制的原因本研究集中于miR-196 a的作用，因此不涉及其他miRNA;因此，在本发明中，研究结果的广泛意义我们的数据揭示了改变的miRNA表达和激活的MEK/ERK激酶，轻微或轻度子宫内膜异位症在位内膜中ERK信号传导我们发现miR-196 a水平与通过MEK/ERK降低的PGR亚型表达相关，表明miR-196 a和MEK/ERK都是子宫内膜异位症的潜在生物标志物。这些结果提供了一种新的方法来靶向子宫内膜异位症孕激素抵抗背后的机制。研究基金/竞争兴趣（S）这项研究是...
STUDY QUESTIONDo microRNAs (miRNAs) contribute to aberrant progesterone receptor (PGR) expression in the eutopic endometrium of women with endometriosis?SUMMARY ANSWERmiR-196a upregulates MEK/ERK signalling, mediating a downregulation of PGR expression in the eutopic endometrium of women with minimal or mild endometriosis.WHAT IS KNOWN ALREADYImplantation failure is strongly suggested as an underlying cause for the observed infertility in minimal or mild endometriosis. Progesterone resistance, which is mainly caused by aberrantly expressed progesterone receptor in the eutopic endometrium, is considered as a key factor of decreased endometrial receptivity; thus far, epigenetics, but not miRNA, has been shown to affect PGR expression in the endometrium.STUDY DESIGN SIZE, DURATIONMicroarray analysis was used to analyse the eutopic endometrium. The differential expression of miR-196a was validated. Bioinformatics analysis predicted that miR-196a targets the 3′-untranslated region (UTR) of the PGR. The relationship between the miR-196a level and PGR expression was studied and the role of the MEK/ERK signal pathway was investigated.PARTICIPANTS/MATERIALS, SETTING, METHODSTotal RNA was extracted from eutopic endometrium samples in three infertile women with mild/minimal endometriosis and three disease-free control subjects. The miRNA and mRNA expression levels were analysed by microarray analysis. The miR-196a expression was validated by qRT-PCR [endometriosis (n= 22) and control (n= 20)], while functional analysis utilisedin vitrotransfection of endometrial stromal cells (ESCs), induction of decidualization of ESCs, and luciferase reporter assays in 293 T cell lines.MAIN RESULTS AND THE ROLE OF CHANCEA total of 66 dysregulated miRNAs and 357 dysregulated mRNAs were screened by microarray analysis. miR-196a and P-MEK/P-ERK were both found to be significantly upregulated in the eutopic endometrium in patients with mild/minimal endometriosis. PGR and PGR-B mRNA were inhibited by miR-196a overexpression and upregulated by miR-196a inhibition. Luciferase reporter failed to confirm the target regulation of miR-196a on PGR. Transfection of ESCs with a miR-196a mimic led to an increase in the P-MEK/P-ERK protein levels, decrease in the PGR protein levels, and atypical decidualization. Following miR-196a inhibition, the P-MEK/P-ERK protein was downregulated and the PGR protein was upregulated. Inhibition of P-MEK/P-ERK also increased PGR expression.LARGE SCALE DATAData are presented in Supplementary Tables SI and SII.LIMITATIONS REASONS FOR CAUTIONThis study focused on the role of miR-196a, and therefore does not involve other miRNAs; hence, it is possible that other miRNAs may also be responsible for progestin resistance in endometriosis.WIDER IMPLICATIONS OF THE FINDINGSOur data revealed altered miRNA expression and activated MEK/ERK signalling in the eutopic endometrium in minimal or mild endometriosis. We showed that the miR-196a level is associated with reduced expression of PGR isoforms through MEK/ERK, suggesting that miR-196a and MEK/ERK are both potential biomarkers of endometriosis. These results provide a novel approach to target the mechanisms behind progesterone resistance in endometriosis.STUDY FUNDING/COMPETING INTEREST(S)This research was …