Role of lncRNA XIST/miR-146a Axis in Matrix Degradation and Apoptosis of Osteoarthritic Chondrocytes Through Regulation of MMP-13 and BCL2.

IF 3.6 3区 生物学 Q1 BIOLOGY
Sara Cheleschi, Nicola Mondanelli, Iole Seccafico, Roberta Corsaro, Elena Moretti, Giulia Collodel, Antonella Fioravanti
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Abstract

Growing evidence demonstrates the critical roles of long non-coding RNAs (lncRNAs) in osteoarthritis (OA) pathogenesis. The lncRNA XIST is one of the most commonly studied; however, its function remains unclear. This study aimed to research the molecular mechanism of XIST in human OA chondrocytes. Cells were transfected with small interfering RNA against XIST or with a microRNA (miR)-146a inhibitor in the presence of interleukin (IL)-1β. Viability was detected using MTT; apoptosis using cytometry; and XIST, miR-146a, B-cell lymphoma (BCL)2, and metalloproteinase (MMP)-13 expression using real-time PCR. The analysis of p50 and p65 nuclear factor (NF)-κB was conducted using PCR and immunofluorescence. Our findings showed that XIST was highly expressed in OA chondrocytes when compared to T/C-28a2 lines. Furthermore, XIST silencing significantly promoted survival and limited apoptosis, with a concomitant over expression of BCL2, reduction in MMP-13 mRNA, and NF-κB activation after IL-1β stimulus. Conversely, miR-146a was significantly down-regulated in OA cells, while its levels were increased following XIST silencing; moreover, miR-146a inhibition induced opposite results to those caused by XIST. Finally, the down-regulation of XIST was correlated to the over-expression of miR-146a, with a consequent modulation of BCL2, MMP-13, and NF-κB. This study suggests an influence of the XIST/miR-146a axis on the viability, apoptosis, and matrix degradation occurring in OA.

lncRNA XIST/miR-146a轴通过调控MMP-13和BCL2在骨关节炎软骨细胞基质降解和凋亡中的作用
越来越多的证据表明,长链非编码rna (lncRNAs)在骨关节炎(OA)发病机制中的关键作用。lncRNA XIST是最常见的研究之一;然而,其功能尚不清楚。本研究旨在探讨XIST在人OA软骨细胞中的分子机制。在白细胞介素(IL)-1β存在的情况下,用靶向XIST的小干扰RNA或microRNA (miR)-146a抑制剂转染细胞。MTT法检测细胞活力;细胞凋亡;以及XIST、miR-146a、b细胞淋巴瘤(BCL)2和金属蛋白酶(MMP)-13的表达。采用PCR和免疫荧光法分析p50和p65核因子(NF)-κB。我们的研究结果表明,与T/C-28a2细胞系相比,XIST在OA软骨细胞中高表达。此外,XIST沉默可显著提高生存率,限制细胞凋亡,同时伴有BCL2的过表达、MMP-13 mRNA的减少和IL-1β刺激后NF-κB的激活。相反,miR-146a在OA细胞中显著下调,而在XIST沉默后其水平升高;此外,miR-146a抑制诱导的结果与XIST相反。最后,XIST的下调与miR-146a的过表达相关,随之而来的是BCL2、MMP-13和NF-κB的调节。本研究表明,XIST/miR-146a轴对OA细胞的活力、凋亡和基质降解有影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biology-Basel
Biology-Basel Biological Science-Biological Science
CiteScore
5.70
自引率
4.80%
发文量
1618
审稿时长
11 weeks
期刊介绍: Biology (ISSN 2079-7737) is an international, peer-reviewed, quick-refereeing open access journal of Biological Science published by MDPI online. It publishes reviews, research papers and communications in all areas of biology and at the interface of related disciplines. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files regarding the full details of the experimental procedure, if unable to be published in a normal way, can be deposited as supplementary material.
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