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首页> 外文期刊>Journal of cellular biochemistry. >Role of miR‐203 in estrogen receptor‐mediated signaling in the rat uterus and endometrial carcinoma
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Role of miR‐203 in estrogen receptor‐mediated signaling in the rat uterus and endometrial carcinoma

机译:miR-203在大鼠子宫和子宫内膜癌中雌激素受体介导的信号传导的作用

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Abstract The role of microRNAs (miRNA) in estrogen receptor (ER) signaling in the uterus and in endometrial cancer is not well understood. We therefore analyzed miRNA expression in uterine samples from a standard 3‐day uterotrophic assay using young female adult rats to identify E2‐regulated miRNAs. Microarray analysis identified 47 E2 down‐regulated miRNAs including miR‐30a, and 25 E2up‐regulated miRNAs including miR‐672, miR‐203, and miR‐146b. The strongly E2‐upregulated miR‐203 was selected for further analysis. miR‐203 was deleted in the rat endometrial adenocarcinoma cell line, RUCA‐I, using CRISPR/CAS9. Five clones devoid of miR‐203 expression were generated. Proliferation was reduced and G2‐arrest was observed in all miR‐203 deficient RUCA‐I clones. Transfection with a miR‐203‐3p mimic partially rescues this effect. Comparison of mRNA expression in three miR‐203 knockout clones to wild type RUCA‐I cells reveals 566 miR‐203‐upregulated and 592 miR‐203‐downregulated genes. 43 of the genes that are upregulated by miR‐203 knockout in vitro are downregulated in the uterus by E2. Of these Acer2 , Zbtb20 , Ptn , Rcbtb2 , Mum1l1 , Hmgn3 , and Nfat5 possess one or more seed sequence matches in their 3′‐UTR that are predicted to be targets of miR‐203. These data demonstrate the importance of E2 regulated miRNAs in general, and miR‐203 in particular, for E2 regulated gene expression and physiological processes including proliferation and cell migration, in the uterus as well as in the etiology of endometrial carcinomas.
机译:摘要microRNA(miRNA)在子宫和子宫内膜癌中雌激素受体(ER)信号传导中的作用尚不清楚。因此,我们使用年轻雌性成年大鼠,通过标准的3天子宫萎缩试验分析子宫样本中的miRNA表达,以确定E2调节的miRNA。微阵列分析确定了47个E2下调的miRNA,包括miR-30a,以及25个E2上调的miRNA,包括miR-672、miR-203和miR-146b。选择E2高度上调的miR-203进行进一步分析。使用CRISPR/CAS9在大鼠子宫内膜腺癌细胞系RUCA-I中删除miR-203。产生了五个没有miR-203表达的克隆。在所有miR-203缺陷型RUCA-I克隆中,增殖减少,G2期阻滞被观察到。用miR-203-3p模拟物转染可部分挽救这种效应。将三个miR-203敲除克隆的mRNA表达与野生型RUCA-I细胞进行比较,发现566个miR-203上调基因和592个miR-203下调基因。43个在体外被miR-203敲除上调的基因在子宫中被E2下调。在这些Acer2、ZBT20、Ptn、Rcbtb2、Mum1l1、Hmgn3和Nfat5中,它们的3′-UTR中有一个或多个种子序列匹配,被预测为miR-203的靶点。这些数据证明了E2调节的miRNA在子宫以及子宫内膜癌病因学中的重要性,尤其是miR-203在E2调节的基因表达和生理过程中的重要性,包括增殖和细胞迁移。

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