Yang Jie, Zhou Yunping, Liang Xiaojun, Jing Bingfei, Zhao Zandong
Department of Foot and Ankle Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an, China.
Department of Hand Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an, China.
Bone Joint Res. 2021 Jul;10(7):459-466. doi: 10.1302/2046-3758.107.BJR-2019-0251.R4.
Osteoarthritis (OA) is characterized by persistent destruction of articular cartilage. It has been found that microRNAs (miRNAs) are closely related to the occurrence and development of OA. The purpose of the present study was to investigate the mechanism of miR-486 in the development and progression of OA.
The expression levels of miR-486 in cartilage were determined by quantitative real-time polymerase chain reaction (qRT-PCR). The expression of collagen, type II, alpha 1 (COL2A1), aggrecan (ACAN), matrix metalloproteinase (MMP)-13, and a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS4) in SW1353 cells at both messenger RNA (mRNA) and protein levels was determined by qRT-PCR, western blot, and enzyme-linked immunosorbent assay (ELISA). Double luciferase reporter gene assay, qRT-PCR, and western blot assay were used to determine whether silencing information regulator 6 (SIRT6) was involved in miR-486 induction of chondrocyte-like cells to a more catabolic phenotype.
Compared with osteonecrosis, the expression of miR-486 was significantly upregulated in cartilage from subjects with severe OA. In addition, overexpressed miR-486 promoted a catabolic phenotype in SW1353 cells by upregulating the expressions of ADAMTS4 and MMP-13 and down-regulating the expressions of COL2A1 and ACAN. Conversely, inhibition of miR-486 had the opposite effect. Furthermore, overexpression of miR-486 significantly inhibited the expression of SIRT6, confirming that SIRT6 is a direct target of miR-486. Moreover, SW1353 cells were transfected with small interfering RNA (si)-SIRT6 and it was found that SIRT6 was involved in and inhibited miR-486-induced changes to SW1353 gene expression.
Our results indicate that miR-486 promotes a catabolic phenotype in SW1353 cells in OA by targeting SIRT6. Our findings might provide a potential therapeutic target and theoretical basis for OA. Cite this article: 2021;10(7):459-466.
骨关节炎（OA）的特征是关节软骨持续破坏。研究发现，微小RNA（miRNA）与OA的发生发展密切相关。本研究旨在探讨miR-486在OA发生发展过程中的作用机制。
采用定量实时聚合酶链反应（qRT-PCR）检测软骨中miR-486的表达水平。运用qRT-PCR、蛋白质印迹法和酶联免疫吸附测定（ELISA）分别检测SW1353细胞中Ⅱ型胶原α1链（COL2A1）、聚集蛋白聚糖（ACAN）、基质金属蛋白酶（MMP）-13和含血小板反应蛋白基序的解聚素和金属蛋白酶-4（ADAMTS4）在信使核糖核酸（mRNA）和蛋白质水平的表达。采用双荧光素酶报告基因检测、qRT-PCR和蛋白质印迹法检测沉默信息调节因子6（SIRT6）是否参与miR-486诱导软骨样细胞向分解代谢表型转变。
与骨坏死相比，严重OA患者软骨中miR-486的表达显著上调。此外，过表达的miR-486通过上调ADAMTS4和MMP-13的表达以及下调COL2A1和ACAN的表达，促进SW1353细胞向分解代谢表型转变。相反，抑制miR-486则产生相反的效果。此外，miR-486的过表达显著抑制SIRT6的表达，证实SIRT6是miR-486的直接靶点。此外，用小干扰RNA（si）-SIRT6转染SW1353细胞，发现SIRT6参与并抑制miR-486诱导的SW1353基因表达变化。
我们的结果表明miR-486通过靶向SIRT6促进OA中SW1353细胞的分解代谢表型。我们的研究结果可能为OA提供潜在的治疗靶点和理论依据。引用本文：2021；10（7）：459-466。
