MicroRNA221-3p modulates Ets-1 expression in synovial fibroblasts from patients with osteoarthritis of temporomandibular joint
MicroRNA221-3p modulates Ets-1 expression in synovial fibroblasts from patients with osteoarthritis of temporomandibular joint
复制标题
MicroRNA221-3p调节颞下颌关节骨关节炎患者滑膜成纤维细胞中Ets-1的表达
DOI：
10.1016/j.joca.2016.06.011
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发表时间：
2016-11-01
影响因子：
7
通讯作者：
Ke, J.
中科院分区：
文献类型：
--
作者：
Xu, J.;Liu, Y.;Ke, J.
目的：本研究旨在筛选颞下颌关节骨关节炎（osteoarthritis of temporomandibular joint，TMJOA）患者滑膜成纤维细胞中差异表达的miRNAs，并探讨其功能。利用实时荧光定量PCR技术对筛选出的miRNA 221 - 3 p进行定量表达，并利用生物信息学方法预测其特异性靶基因。将miRNA 221 - 3 p模拟物或抑制剂转染滑膜成纤维细胞后，分别用免疫组化、实时荧光PCR和Western blot检测v-Ets禽成红细胞增多症病毒E26癌基因同源物1（Ets-1）的表达。进行双荧光素酶活性以鉴定miRNA 221 - 3 p对Ets-1的直接调节。结果：在TMJOA滑膜成纤维细胞中，有8个miRNAs表达上调，6个miRNAs表达下调。MiRNA 221 - 3 p的表达最低。Ets-1的3 '-非翻译（3'-UTR）序列，与miRNA 221 - 3 p的种子序列互补。Ets-1的高表达与miRNA 221 - 3 p的减弱相关。在TMJOA滑膜成纤维细胞中，miRNA 221 - 3 p的过表达抑制了Ets-1转录本3 '-UTR的报告基因的活性，并抑制了Ets-1及其下游分子基质金属蛋白酶1（MMP 1）和MMP 9的表达。结论：TMJOA患者滑膜成纤维细胞中miRNA 221 - 3 p表达减少，可能与炎症环境中IL-1 β的大量分泌有关，这可能通过诱导Ets-1的表达上调，进而启动MMP 1和MMP 9的分泌，导致TMJOA的持续病理发展。(C)2016国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: This study aimed to screen differential expression of microRNAs ( miRNAs), and investigate function of the specifically selected miRNA in synovial fibroblasts from patients suffering osteoarthritis of temporomandibular joint ( TMJOA).Methods: MiRNA microarray was used to select differentially expressed miRNAs between TMJOA and normal synovial fibroblasts. The expression of screened miRNA221-3p was quantified using real-time PCR, and its specific target gene was predicted by bioinformatics. After transfection of miRNA221-3p mimics or inhibitor into synovial fibroblasts, the expression of v-Ets avian erythroblastosis virus E26 oncogene homolog 1 ( Ets-1) was detected by immunohistochemistry, real-time PCR and Western blot, respectively. Dual luciferase activity was performed to identify the direct regulation of miRNA221-3p on Ets-1. Interlukin-1 beta ( IL-1 beta) mimics an inflammatory situation.Results: In TMJOA synovial fibroblasts, eight miRNAs were up-regulated and six miRNAs were down-regulated. MiRNA221-3p was the most down-expressed. A sequence in the 3'-untranslated ( 3'-UTR) of Ets-1 complementary to the seed sequence of miRNA221-3p. Elevated expression of Ets-1 associated with attenuation of miRNA221-3p. Over-expression of miRNA221-3p suppressed the activity of a reporter construct containing the 3'-UTR of Ets-1 transcript and inhibited the expression of Ets-1 as well as its downstream molecules, matrix metalloproteinase 1 ( MMP1) and MMP9 in TMJOA synovial fibroblasts. IL-1 beta suppressed the expression of miRNA221-3p in both a dose-dependent and time-dependent manner.Conclusion: The reduction of miRNA221-3p in synovial fibroblasts, attributed from abundance of IL-1 beta in inflamed circumstance, induces Ets-1 up-regulation and then, initiates MMP1 and MMP9 secretion, thereby leading to continuously pathological development in TMJOA. (C) 2016 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.