Open AccessMicroRNA-486 promotes a more catabolic phenotype in chondrocyte-like cells by targeting SIRT6
Author(s) -
Jie Yang,
Yue Zhou,
Xiaojun Liang,
Bingfei Jing,
Zandong Zhao
Publication year - 2021
Publication title -
bone and joint research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.639
H-Index - 31
ISSN - 2046-3758
DOI - 10.1302/2046-3758.107.bjr-2019-0251.r4
Subject(s) - aggrecan , chondrocyte , adamts , gene silencing , small interfering rna , microrna , western blot , transfection , metalloproteinase , downregulation and upregulation , microbiology and biotechnology , gene knockdown , chemistry , biology , thrombospondin , matrix metalloproteinase , cartilage , cell culture , gene , osteoarthritis , medicine , genetics , pathology , biochemistry , articular cartilage , alternative medicine , anatomy
Aims 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.Methods 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.Results 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.Conclusion 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: Bone Joint Res 2021;10(7):459–466.