Mutual regulation between microRNA-373 and methyl-CpG-binding domain protein 2 in hilar cholangiocarcinoma
Mutual regulation between microRNA-373 and methyl-CpG-binding domain protein 2 in hilar cholangiocarcinoma
复制标题
microRNA-373与甲基CpG结合域蛋白2在肝门部胆管癌中的相互调节
DOI：
10.3748/wjg.v18.i29.3849
复制
发表时间：
2012-08-07
影响因子：
4.3
通讯作者：
Zou, Sheng-Quan
中科院分区：
文献类型：
--
作者：
Chen, Yong-Jun;Luo, Jian;Zou, Sheng-Quan
目的：通过探讨miR-373与甲基化CpG结合域蛋白（methyl-CpG-binding domain protein，MBD）2之间的相关性，探讨microRNA（microRNA，miRNA）与DNA甲基化之间的相互调节作用。使用甲基化特异性聚合酶链反应研究miR-373的甲基化，使用染色质免疫沉淀测定研究甲基结合蛋白的募集。使用QuikChangeTM定点诱变试剂盒进行突变分析。结果：在肝门部胆管癌中，miR-373表达减少，且与细胞分化差、临床分期晚、生存期短密切相关。miR-373基因启动子相关的CpG岛甲基化，抑制miR-373的表达。MBD 2在miR-373的启动子相关CpG岛上调并富集。结论：miR-373是MBD 2的直接转录靶点，通过CpG岛甲基化的反馈环负调控MBD 2的活性。(c)2012年百世登。All rights reserved.
AIM: To investigate the reciprocal modulation between microRNA (miRNA) and DNA methylation via exploring the correlation between miR-373 and methyl-CpG-binding domain protein (MBD)2.METHODS: MiR-373 expression was examined using the TaqMan miRNA assay. Methylation of miR-373 was investigated using methylation-specific polymerase chain reaction, and recruitment of methyl binding proteins was studied using the chromatin immunoprecipitation assay. Mutation analysis was conducted using the QuikChange (TM) Site-Directed Mutagenesis kit. The activity of miR-373 gene promoter constructs and targeting at MBD2-three prime untranslated region (3'UTR) by miR-373 were evaluated by a dual-luciferase reporter gene assay.RESULTS: In hilar cholangiocarcinoma, miR-373 decreased and was closely associated with poor cell differentiation, advanced clinical stage, and shorter survival. The promoter-associated CpG island of miR-373 gene was hypermethylated and inhibited expression of miR-373. MBD2 was up-regulated and enriched at the promoter-associated CpG island of miR-373. Methylation-mediated suppression of miR-373 required MBD2 enrichment at the promoter-associated CpG island, and miR-373 negatively regulated MBD2 expression through targeting the 3'UTR.CONCLUSION: MiR-373 behaves as a direct transcriptional target and negative regulator of MBD2 activity through a feedback loop of CpG island methylation. (c) 2012 Baishideng. All rights reserved.