miR-30-HNF4γ and miR-194-NR2F2 regulatory networks contribute to the upregulation of metaplasia markers in the stomach.
miR-30-HNF4γ and miR-194-NR2F2 regulatory networks contribute to the upregulation of metaplasia markers in the stomach.
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DOI：
10.1136/gutjnl-2014-308759
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发表时间：
2016-06
期刊：
影响因子：
24.5
通讯作者：
Goldenring JR
中科院分区：
文献类型：
--
作者：
Sousa JF;Nam KT;Petersen CP;Lee HJ;Yang HK;Kim WH;Goldenring JR
肠化生和痉挛性多肽表达化生（SPEM）被认为是胃腺癌的肿瘤前体，与正常胃相比，两者都以基因表达改变为标志。由于miRNAs是基因表达的重要调节因子，我们试图研究miRNAs在胃化生发展中的作用。我们使用qRT-PCR方法对激光捕获显微切割的人肠上皮化生和SPEM进行miRNA分析。进行miRNA谱与来自相同样品的先前mRNA谱的数据整合以检测潜在的miRNA-mRNA调控回路。用选定的miRNA模拟物和抑制剂转染胃癌细胞系，以评估它们对推定靶点和其他化生标志物表达的影响。我们确定了几个基因作为在化生进展过程中改变的miRNA的潜在靶点。我们证明了HNF 4 γ（在肠化生中上调）被miR-30靶向，而miR-194靶向HNF 4活性的已知共调节因子NR 2F 2（在肠化生中下调）。miR-30 a过表达后，肠上皮化生标志物VIL 1、TFF 2和TFF 3以HNF 4 γ依赖的方式下调。此外，HNF 4 γ的过表达足以诱导VIL 1的表达，并且这种作用通过下调NR 2F 2而增强。两种转录因子HNF 4 γ和NR 2F 2的相互作用以及它们分别由miR-30和miR-194的协调调节代表了在肠上皮化生发展期间负责胃细胞谱系中肠转录物表达的miRNA至转录因子网络。
Intestinal metaplasia and spasmolytic polypeptide-expressing metaplasia (SPEM) are considered neoplastic precursors of gastric adenocarcinoma and are both marked by gene expression alterations in comparison to normal stomach. Since miRNAs are important regulators of gene expression, we sought to investigate the role of miRNAs on the development of stomach metaplasias. We performed miRNA profiling using a qRT-PCR approach on laser capture microdissected human intestinal metaplasia and SPEM. Data integration of the miRNA profile with a previous mRNA profile from the same samples was performed to detect potential miRNA-mRNA regulatory circuits. Transfection of gastric cancer cell lines with selected miRNA mimics and inhibitors was used to evaluate their effects on the expression of putative targets and additional metaplasia markers. We identified several genes as potential targets of miRNAs altered during metaplasia progression. We showed evidence that HNF4γ (upregulated in intestinal metaplasia) is targeted by miR-30 and that miR-194 targets a known co-regulator of HNF4 activity, NR2F2 (downregulated in intestinal metaplasia). Intestinal metaplasia markers such as VIL1, TFF2 and TFF3 were down-regulated after overexpression of miR-30a in a HNF4γ-dependent manner. In addition, overexpression of HNF4γ was sufficient to induce the expression of VIL1 and this effect was potentiated by down-regulation of NR2F2. The interplay of the two transcription factors HNF4γ and NR2F2 and their coordinate regulation by miR-30 and miR-194, respectively, represent a miRNA to transcription factor network responsible for the expression of intestinal transcripts in stomach cell lineages during the development of intestinal metaplasia.