Department of Orthopedics, Xi'an Fifth Hospital, Xi'an, Shaanxi, PR China.
Department of Orthopedics, Shaanxi Provincial People's Hospital, Xian City, Shaanxi, PR China.
Bioengineered. 2022 Apr;13(4):11137-11145. doi: 10.1080/21655979.2022.2061287.
It is known that long intergenic non-protein coding RNA 00707 (LINC00707) promotes lipopolysaccharide (LPS)-injury and microRNA-199-3p (miR-199-3p) regulates chondrocyte proliferation and apoptosis. Both processes participate in osteoarthritis (OA). We predicted that LINC00707 and miR-199-3p may interact with each other. Therefore, LINC00707 and miR-199-3p may interact with each other to participate in OA. In this study, the expression of LINC00707 and miR-199-3p in both OA and normal articular cartilage tissues was analyzed using RT-qPCR. The subcellular location of LINC00707 and its direct interaction with miR-199-3p were explored by nuclear fractionation assay, RNA pull-down assay and Luciferase reporter assay, respectively. The role of LINC00707 and miR-199-3p in regulating the expression of each other was analyzed using an overexpression assay, followed by RT-qPCR. The role of LINC00707 and miR-199-3p in regulating OA chondrocyte proliferation and apoptosis was analyzed by BrdU assay and cell apoptosis assay, respectively. OA tissues exhibited increased expression of LINC00707 and decreased expression of miR-199-3p. LINC00707 directly interacted with miR-199-3p in cytoplasm. However, LINC00707 and miR-199-3p overexpression failed to affect each other's expression. LPS treatment increased LINC00707 expression and decreased miR-199-3p expression in OA chondrocyte. LINC00707 overexpression increased the apoptosis of OA chondrocytes induced by LPS and suppressed the proliferation of OA chondrocytes. Moreover, LINC00707 suppressed the role of miR-199-3p in enhancing cell proliferation and suppressing cell apoptosis. In conclusion, LINC00707 can be detected in cytoplasm and it may sponge miR-199-3p to regulate chondrocyte proliferation and apoptosis in OA.
已知长链非编码 RNA 00707(LINC00707)可促进脂多糖(LPS)损伤,微小 RNA-199-3p(miR-199-3p)调控软骨细胞增殖和凋亡,这两个过程均参与骨关节炎(OA)。我们预测 LINC00707 和 miR-199-3p 可能相互作用。因此,LINC00707 和 miR-199-3p 可能通过相互作用参与 OA。在这项研究中,通过 RT-qPCR 分析 OA 和正常关节软骨组织中 LINC00707 和 miR-199-3p 的表达。通过核分馏测定、RNA 下拉测定和荧光素酶报告基因测定分别探讨 LINC00707 的亚细胞定位及其与 miR-199-3p 的直接相互作用。通过过表达测定分析 LINC00707 和 miR-199-3p 调控彼此表达的作用,然后通过 RT-qPCR 进行分析。通过 BrdU 测定和细胞凋亡测定分别分析 LINC00707 和 miR-199-3p 对 OA 软骨细胞增殖和凋亡的调节作用。OA 组织中 LINC00707 表达增加,miR-199-3p 表达降低。LINC00707 与细胞质中的 miR-199-3p 直接相互作用。然而,LINC00707 和 miR-199-3p 的过表达未能影响彼此的表达。LPS 处理增加 OA 软骨细胞中 LINC00707 的表达并降低 miR-199-3p 的表达。LINC00707 过表达增加 LPS 诱导的 OA 软骨细胞凋亡并抑制 OA 软骨细胞增殖。此外,LINC00707 抑制 miR-199-3p 增强细胞增殖和抑制细胞凋亡的作用。总之,LINC00707 可在细胞质中检测到,它可能通过海绵吸附 miR-199-3p 来调节 OA 中的软骨细胞增殖和凋亡。