MicroRNA miR-466 inhibits Lymphangiogenesis by targeting prospero-related homeobox 1 in the alkali burn corneal injury model.
MicroRNA miR-466 inhibits Lymphangiogenesis by targeting prospero-related homeobox 1 in the alkali burn corneal injury model.
复制标题
DOI：
10.1186/s12929-014-0104-0
复制
发表时间：
2015-01-02
影响因子：
11
通讯作者：
Lee SK
中科院分区：
文献类型：
--
作者：
Seo M;Choi JS;Rho CR;Joo CK;Lee SK
淋巴管生成是角膜移植排斥反应的主要原因之一。在淋巴管生成因子中，血管内皮生长因子（VEGF）-C和-D被认为是最有效的。两者均与VEGF受体3（VEGFR 3）结合以激活Prospero同源框1（Prox 1），这是一种对淋巴管系统的发育和维持至关重要的转录因子。微小RNA（miRNAs）以序列特异性方式结合靶基因的3'非翻译区（3' UTR）并抑制基因表达。在目前的研究中，我们寻找靶向促淋巴管生成因子Prox 1的miRNAs。在通过生物信息学分析预测的与Prox-1的3' UTR种子匹配的miRNA中，我们选择了3个（miR-466、miR-4305和miR-4795- 5 p）用于进一步研究。miR-466和miR-4305模拟物，但不是miR-4795- 5 p模拟物，显著降低了Prox-1 3' UTR报告载体的荧光素酶活性。在原代淋巴管内皮细胞（HDLEC）中，miR-466模拟转染抑制Prox 1 mRNA和蛋白表达，而miR-4305模拟转染则没有。使用Prox 1的3' UTR上的两个可能的种子匹配位点的突变的报告构建体的实验表明靶位点2直接结合miR-466。与乱序对照相比，用miR-466模拟物转染的HDLEC抑制管形成。此外，与对照抑制剂转染的细胞相比，用miR-466抑制剂转染的HDLEC显示出增强的管形成，并且这种抑制作用被Prox 1 siRNA抵消。与乱序对照相比，miR-466模拟物减少了血管生成和淋巴管生成，导致角膜损伤大鼠模型中的角膜更清晰。我们的数据表明，miR-446可能通过在转录后水平抑制Prox 1表达对移植角膜具有保护作用。目前的研究结果可能会提供深入了解角膜移植排斥反应和碱烧伤造成的淋巴管生成的机制，以及淋巴管生成性眼病的新治疗方法的发展。本文的在线版本（doi：10.1186/s12929-014-0104-0）包含补充材料，可供授权用户使用。
Lymphangiogenesis is one of the major causes of corneal graft rejection. Among the lymphangiogenic factors, vascular endothelial growth factor (VEGF)-C and -D are considered to be the most potent. Both bind to VEGF receptor 3 (VEGFR3) to activate Prospero homeobox 1 (Prox1), a transcription factor essential for the development and maintenance of lymphatic vasculature. MicroRNAs (miRNAs) bind to the 3' untranslated regions (3' UTRs) of target genes in a sequence-specific manner and suppress gene expression. In the current study, we searched for miRNAs that target the pro-lymphangiogenic factor Prox1. Among the miRNAs predicted by the bioinformatic analysis to seed match with the 3' UTR of Prox-1, we chose 3 (miR-466, miR-4305, and miR-4795-5p) for further investigation. Both the miR-466 and miR-4305 mimics, but not the miR-4795-5p mimic, significantly reduced the luciferase activity of the Prox-1 3' UTR reporter vector. In primary lymphatic endothelial cells (HDLEC), miR-466 mimic transfection suppressed Prox1 mRNA and protein expression, while miR-4305 mimic transfection did not. Experiments using mutated reporter constructs of the two possible seed match sites on the 3' UTR of Prox1 suggested that the target site 2 directly bound miR-466. HDLEC transfected with the miR-466 mimic suppressed tube formation as compared to the scrambled control. Furthermore, HDLEC transfected with a miR-466 inhibitor showed enhanced tube formation as compared to control inhibitor transfected cells, and this inhibitory effect was counteracted by Prox1 siRNA. The miR-466 mimic reduced angiogenesis and lymphangiogenesis resulting in clearer corneas in an cornea injury rat model compared to the scrambled control. Our data suggest that miR-446 may have a protective effect on transplanted corneas by suppressing Prox1 expression at the post-transcriptional level. The results of the current study may provide insights into the mechanisms of lymphangiogenesis resulting from corneal graft rejection and alkali-burn injuries, as well as into the development of new treatments for lymphangiogenic eye diseases. The online version of this article (doi:10.1186/s12929-014-0104-0) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
2.8
作者：
Cursiefen, C;Chen, L;Streilein, JW
通讯作者：
Streilein, JW
影响因子：
45.3
作者：
Giantonio, Bruce J.;Catalano, Paul J.;Benson, Al B., III
通讯作者：
Benson, Al B., III
DOI：
10.1073/pnas.95.24.14389
发表时间：
1998-11-24
影响因子：
11.1
作者：
Cao, YH;Linden, P;Alitalo, K
通讯作者：
Alitalo, K
DOI：
10.4049/jimmunol.0903180
发表时间：
2010-01-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Dietrich T;Bock F;Yuen D;Hos D;Bachmann BO;Zahn G;Wiegand S;Chen L;Cursiefen C
通讯作者：
Cursiefen C
DOI：
10.1073/pnas.95.2.548
发表时间：
1998-01-20
影响因子：
11.1
作者：
Achen, MG;Jeltsch, M;Stacker, SA
通讯作者：
Stacker, SA