Department of Orthopaedics, Fuyang People's Hospital, Fuyang, China.
Clinical Medical College of Six Hospitals, Shanghai Jiao Tong University, Shanghai, China.
J Gene Med. 2020 Sep;22(9):e3203. doi: 10.1002/jgm.3203. Epub 2020 May 1.
Non-coding RNAs are endogenous regulators of gene expression that have been implicated in the pathogenesis of various diseases, including osteoarthritis (OA). Long non-coding RNA nuclear enriched abundant transcript 1 (NEAT1) and miR-16-5p are up-regulated in OA tissues; however, their functions have not been clarified.
Chondrocyte ATDC5 was used as a cell model. NEAT1 overexpression and knockdown cells were established by transfection with lipofectamine. miR-16-5p was also transfected into the cells using lipofectamine. Moreover, cell proliferation was examined using cell counting kit-8 assays. Cell apoptosis was evaluated by flow cytometry. The interaction between NEAT1 and miR-16-5p was validated by a Quantitative real-time RT-PCR (qRT-PCR) and dual-luciferase reporter assays.
NEAT1 could increase cell viability and decrease apoptosis of ATDC5 cells, whereas miR-16-5p had the opposite effects. NEAT1 could specifically bind to miR-16-5p and reduce its expression.
The suppression of miR-16-5p, as mediated by NEAT1 overexpression, could promote proliferation and inhibit apoptosis of chondrocytes. It was also revealed that NEAT1 is a "double-edged sword" during the development of OA.
非编码 RNA 是基因表达的内源性调节剂，它们与各种疾病的发病机制有关，包括骨关节炎（OA）。长链非编码 RNA 核富集丰富转录物 1（NEAT1）和 miR-16-5p 在 OA 组织中上调；然而，它们的功能尚未阐明。
使用软骨细胞 ATDC5 作为细胞模型。通过脂质体转染建立 NEAT1 过表达和敲低细胞。使用脂质体将 miR-16-5p 转染到细胞中。此外，通过细胞计数试剂盒-8 测定法检查细胞增殖。通过流式细胞术评估细胞凋亡。通过定量实时 RT-PCR（qRT-PCR）和双荧光素酶报告基因测定验证 NEAT1 和 miR-16-5p 之间的相互作用。
NEAT1 可以增加 ATDC5 细胞的活力并减少细胞凋亡，而 miR-16-5p 则具有相反的作用。NEAT1 可以特异性结合 miR-16-5p 并降低其表达。
通过过表达 NEAT1 抑制 miR-16-5p 可以促进软骨细胞的增殖并抑制细胞凋亡。研究还表明，NEAT1 在 OA 的发展过程中是一把“双刃剑”。
