BACKGROUND This study aimed to investigate the role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) and related molecular mechanisms, in osteoarthritis (OA). METHODS Cartilage tissues of OA patients and healthy volunteers were isolated and cultured. After transfection with the appropriate constructs, chondrocytes were classified into Blank, pcDNA3.1-NC, pcDNA3.1-MEG3, si-NC, si-MEG3, pcDNA3.1-NC + mimics NC, pcDNA3.1-MEG3 + mimics NC, pcDNA3.1-NC + miR-361-5p mimics and pcDNA3.1-MEG3 + miR-361-5p mimics groups. qRT-PCR was used to detect the expression of MEG3, miR-361-5p and FOXO1. Western blot, luciferase reporter assay, RIP, CCK-8, and flow cytometry analysis were performed to reveal the morphology, proliferation, and apoptotic status of cartilage cells. Histological analysis and immunostaining were conducted in the OA rat model. RESULTS Expression of MEG3 and FOXO1 was significantly decreased in OA compared with the normal group, while the expression of miR-361-5p was increased. MEG3 might serve as a ceRNA of miR-361-5p in OA chondrocytes. Moreover, using western blot analyses and the CCK-8 assay, MEG3 was shown to target miR-361-5p/FOXO1, elevate cell proliferation, and impair cell apoptosis. Functional analysis in vivo showed that MEG3 suppressed degradation of the cartilage matrix. CONCLUSION MEG3 can contribute to cell proliferation and inhibit cell apoptosis and degradation of extracellular matrix (ECM) via the miR-361-5p/FOXO1 axis in OA chondrocytes.

中文翻译：

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MEG3通过miR-361-5p / FOXO1轴促进骨关节炎软骨细胞的增殖并抑制其凋亡。

背景技术本研究旨在研究母体表达的长非编码RNA（lncRNA）3（MEG3）及其相关分子机制在骨关节炎（OA）中的作用。方法分离培养OA患者和健康志愿者的软骨组织。用适当的构建体转染后，将软骨细胞分类为空白，pcDNA3.1-NC，pcDNA3.1-MEG3，si-NC，si-MEG3，pcDNA3.1-NC +模拟NC，pcDNA3.1-MEG3 +模拟NC ，pcDNA3.1-NC + miR-361-5p模拟物和pcDNA3.1-MEG3 + miR-361-5p模拟物组。用qRT-PCR检测MEG3，miR-361-5p和FOXO1的表达。进行了蛋白质印迹，荧光素酶报告基因分析，RIP，CCK-8和流式细胞仪分析，以揭示软骨细胞的形态，增殖和凋亡状态。在OA大鼠模型中进行组织学分析和免疫染色。结果与正常组相比，OA中MEG3和FOXO1的表达明显降低，而miR-361-5p的表达升高。MEG3可能作为OA软骨细胞中miR-361-5p的ceRNA。此外，使用蛋白质印迹分析和CCK-8分析，MEG3被证明靶向miR-361-5p / FOXO1，提高细胞增殖，并损害细胞凋亡。体内功能分析表明，MEG3抑制了软骨基质的降解。结论MEG3可以通过miR-361-5p / FOXO1轴促进OA软骨细胞的增殖，并抑制细胞凋亡和细胞外基质（ECM）降解。此外，使用蛋白质印迹分析和CCK-8分析，MEG3被证明靶向miR-361-5p / FOXO1，提高细胞增殖，并损害细胞凋亡。体内功能分析表明，MEG3抑制了软骨基质的降解。结论MEG3可以通过miR-361-5p / FOXO1轴促进OA软骨细胞的增殖，并抑制细胞凋亡和细胞外基质（ECM）降解。此外，使用蛋白质印迹分析和CCK-8分析，MEG3被证明靶向miR-361-5p / FOXO1，提高细胞增殖，并损害细胞凋亡。体内功能分析表明，MEG3抑制了软骨基质的降解。结论MEG3可以通过miR-361-5p / FOXO1轴促进OA软骨细胞的增殖，并抑制细胞凋亡和细胞外基质（ECM）降解。