MicroRNA-219c-5p regulates bladder fibrosis by targeting FN1.
MicroRNA-219c-5p regulates bladder fibrosis by targeting FN1.
复制标题
DOI：
10.1186/s12894-020-00765-5
复制
发表时间：
2020-12-07
期刊：
影响因子：
2
通讯作者：
Yang J
中科院分区：
文献类型：
--
作者：
Liu B;Ding Y;Li P;Wang T;He S;Jia Z;Yang J
我们发现多发性硬化症小鼠的膀胱有明显的纤维化。本研究旨在探讨纤维连接蛋白1(FN1)与膀胱纤维化的关系，以及参与FN1调控的microRNAs。免疫组织化学方法观察大鼠膀胱平滑肌纤维化程度。此外，我们使用实时定量聚合酶链式反应(RT-qPCR)和Western blotting来检测FN1在不同程度纤维化的膀胱中的表达。生物信息学分析表明miR-199A-3P、miR-219C-5P和miR-3572-3P可抑制FN1的合成。因此，miR-199a-3p、miR-219C-5p和miR-3572-3p在膀胱平滑肌细胞(BSMCs)中过表达或下调，并通过RT-qPCR检测各自的转染率和FN1基因敲除效率。只有miR-219C-5p过表达和基因敲除产生了预期的结果。用双荧光素酶报告基因分析确定miR-219C-5p与FN1之间的靶向关系。流式细胞仪和细胞计数试剂盒8(CCK8)实验证实，miR-219C-5p降低了FN1的表达，并影响了平滑肌细胞的生物学活性。体内分别将miR-219C-5P的Agomir和Anagomir基因导入小鼠体内，观察其对小鼠膀胱纤维化的影响。随着膀胱纤维化程度的加重，FN1的表达增加，miR-199a-3p、miR-219C-5p和miR-3572-3p的表达水平降低。RT-qPCR结果显示，只有miR-219C-5p对FN1有调节作用。事实上，双荧光素酶报告实验结果表明，miR-219C-5p直接靶向FN1。CCK8和细胞周期分析显示miR-219C-5p过表达抑制BSMC增殖，miR-219C-5p基因敲除促进BSMC增殖。细胞凋亡实验显示，miR-219C-5p过表达促进细胞凋亡，而miR-219C-5p基因敲除抑制BSMC凋亡。体内转染miR-219C-5p的Anagomir和Anagomir发现，Anagomir组小鼠膀胱纤维化减轻，Anagomir组更严重。提示FN1上调和miR-219C-5p下调在膀胱纤维化的发生发展中起重要作用，miR-219C-5p通过调节FN1的表达参与了膀胱纤维化的发生。因此，我们提出了miR-219C-5P的一种新的抗纤维化作用，可能成为诊断和治疗膀胱纤维化的潜在靶点。
We found that the bladders of multiple sclerosis mice were significantly fibrotic. This study aimed to investigate the relationship between fibronectin 1 (FN1) and bladder fibrosis, as well as the microRNAs involved in FN1 regulation. The degree of bladder smooth muscle fibrosis was observed by immunohistochemistry. In addition, we used quantitative real-time polymerase chain reaction (RT-qPCR) and Western blotting to determine FN1 expression in bladders with different grades of fibrosis. Bioinformatics analysis revealed that miR-199a-3p, miR-219c-5p and miR-3572-3p could inhibit FN1 synthesis. Therefore, miR-199a-3p, miR-219c-5p and miR-3572-3p were overexpressed or knocked down in bladder smooth muscle cells (BSMCs), and the respective transfection and FN1 knockdown efficiencies were detected by RT-qPCR. Only miR-219c-5p overexpression and knockdown produced the expected results. A dual luciferase reporter assay was used to determine the targeting relationship between miR-219c-5p and FN1. Flow cytometry and Cell Counting Kit 8 (CCK8) experiments confirmed that miR-219c-5p reduced FN1 expression and affected the biological activity of smooth muscle cells. Agomir and anagomir of miR-219c-5p were transfected in vivo to observe the change of bladder fibrosis in mice. With increasing bladder fibrosis, FN1 expression increased, while miR-199a-3p, miR-219c-5p, and miR-3572-3p expression levels decreased. The RT-qPCR results after transfection showed that only miR-219c-5p could regulate FN1. Indeed, the dual luciferase reporter assay results indicated that miR-219c-5p targeted FN1 directly. CCK8 and cell cycle assays showed that miR-219c-5p overexpression inhibited BSMC proliferation, while miR-219c-5p knockdown promoted BSMC proliferation. An apoptosis assay showed that miR-219c-5p overexpression promoted apoptosis, while miR-219c-5p knockdown inhibited BSMC apoptosis. The agomir and anagomir transfected with miR-219c-5p in vivo found that the bladder fibrosis of the mice in the agomir group was reduced, and the anagomir group was worse. Our findings indicate that FN1 up-regulation and miR-219c-5p down-regulation play an important role in the development of bladder fibrosis, and miR-219c-5p participates in bladder fibrosis by regulating FN1 expression. Thus, a novel antifibrotic function of miR-219c-5p is proposed, which may represent a potential target for the diagnosis and treatment of bladder fibrosis.
登录
查看更多内容
纤连蛋白1通过抑制细胞凋亡和调节EMT促进黑色素瘤增殖和转移
影响因子：
4
作者：
Li, Bifei;Shen, Weiyu;Jia, Lee
通讯作者：
Jia, Lee
DOI：
10.1177/1352458509104594
发表时间：
2009-07-01
期刊：
MULTIPLE SCLEROSIS
影响因子：
--
作者：
Al-Izki, S.;Pryce, G.;Baker, D.
通讯作者：
Baker, D.
影响因子：
16.8
作者：
Steinman, L;Zamvil, SS
通讯作者：
Zamvil, SS
DOI：
10.1152/ajprenal.00273.2012
发表时间：
2012-11-01
影响因子：
4.2
作者：
Altuntas, Cengiz Z.;Daneshgari, Firouz;Tuohy, Vincent K.
通讯作者：
Tuohy, Vincent K.
影响因子：
3.3
作者：
Altuntas CZ;Daneshgari F;Liu G;Fabiyi A;Kavran M;Johnson JM;Gulen MF;Jaini R;Li X;Frenkl TL;Tuohy VK
通讯作者：
Tuohy VK