Knockdown of PVT1 inhibits IL-1β-induced injury in chondrocytes by regulating miR-27b-3p/TRAF3 axis
Knockdown of PVT1 inhibits IL-1β-induced injury in chondrocytes by regulating miR-27b-3p/TRAF3 axis
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DOI：
10.1016/j.intimp.2019.106052
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发表时间：
2020-02-01
影响因子：
5.6
通讯作者：
Yu, Huimin
中科院分区：
文献类型：
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作者：
Lu, Xiuyun;Yu, Yanhui;Yu, Huimin
长非编码 RNA 浆细胞瘤变体易位 1 (PVT1) 已被确定与骨关节炎 (OA) 的进展有关。然而，PVT1 在 OA 发展中的潜在机制仍然很大程度上未知。本研究旨在探讨PVT1对白细胞介素1β（IL-1β）诱导的软骨细胞损伤的影响并探讨潜在机制。收集 25 名 OA 患者和正常对照的软骨组织。 IL-1β刺激人转化软骨细胞C28/I2。通过实时定量聚合酶链反应或蛋白质印迹检测PVT1、微小RNA（miR）-27b-3p和肿瘤坏死因子受体相关因子3（TRAF3）的水平。分别使用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴化物、流式细胞术、蛋白质印迹和酶联免疫吸附测定，通过细胞活力、细胞凋亡、自噬和炎症反应来研究 IL-1 β 诱导的损伤。通过荧光素酶报告基因、RNA 免疫沉淀和 RNA Pull-down 测定探索 miR-27b-3p 与 PVT1 或 TRAF3 之间的靶标关联。我们发现 OA 患者和 IL-1A 处理的 C28/I2 细胞中 PVT1 表达增强。在IL-1β处理的C28/I2细胞中，PVT1的沉默促进了细胞活力和自噬，但抑制了细胞凋亡和炎症反应。 miR-27b-3p 被证实是 PVT1 的靶标，其缺陷逆转了 PVT1 敲低对 IL-1 β 诱导的损伤的抑制作用。 TRAF3 是 miR-27b-3p 的靶标，可减弱 miR-27b-3p 对 IL-1 β 诱导的 C28/I2 细胞损伤的影响。此外，PVT1 通过海绵 miR-27b-3p 正向调节 TRAF3 表达。总的来说，PVT1 的敲低增加了细胞活力和自噬，但通过上调 miR-27b-3p 和下调 TRAF3 来抑制 IL-1 beta 处理的软骨细胞的凋亡和炎症反应。
Long noncoding RNA plasmacytoma variant translocation 1 (PVT1) has been identified to implicate in the progression of osteoarthritis (OA). However, the mechanism underlying PVT1 in OA development remains largely unknown. This study aimed to investigate the effect of PVT1 on interleukin-1 beta (IL-1 beta)-induced injury in chondrocytes and explore potential mechanism. The cartilage tissues from 25 OA patients and normal controls were collected. Human transformed chondrocytes C28/I2 were stimulated by IL-1 beta. The levels of PVT1, microRNA (miR)-27b-3p, and tumor necrosis factor receptor-associated factor 3 (TRAF3) were detected by quantitative real-time polymerase chain reaction or western blot. IL-1 beta-induced injury was investigated by cell viability, apoptosis, autophagy and inflammatory response using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, flow cytometry, western blot and enzyme linked immunosorbent assay, respectively. The target association between miR-27b-3p and PVT1 or TRAF3 was explored by luciferase reporter, RNA immunoprecipitation and RNA pull-down assays. We found that PVT1 expression was enhanced in OA patients and IL-1A-treated C28/I2 cells. Silence of PVT1 promoted cell viability and autophagy but suppressed apoptosis and inflammatory response in IL-1 beta-treated C28/I2 cells. miR-27b-3p was confirmed as a target of PVT1 and its deficiency reversed the suppressive effect of PVT1 knockdown on IL-1 beta-induced injury. TRAF3 was a target of miR-27b-3p and attenuated the effect of miR-27b-3p on IL-1 beta-induced injury in C28/I2 cells. Moreover, TRAF3 expression was positively regulated by PVT1 via sponging miR-27b-3p. Collectively, knockdown of PVT1 increased cell viability and autophagy but inhibited apoptosis and inflammatory response in chondrocytes treated by IL-1 beta via up-regulating miR-27b-3p and down-regulating TRAF3.