长链非编码RNA HOX转录反义RNA对微小RNA-519c-3p/骨形成蛋白Ⅱ型受体调控骨关节炎进展作用及分子机制  

Long noncoding RNA HOX transcript antisense RNA regulates the progression of osteoarthritis via miR-519c-3p/bone morphogenetic protein receptor 2 axis

作　　者：范忠诚[1] 王琮仁[1] 符策岗 陈晓峰 FAN Zhong-cheng;WANG Cong-ren;FU Ce-gang;CHEN Xiao-feng(Department of Orthopedic Center,Haikou Affliated Hospital of Central South University Xiangya School of Medicine,Haikou 570208,China;Department of Orthopedic Center,Haikou Orthopedics and Diabetes Hospital,Haikou 570208,China)

机构地区：[1]中南大学湘雅医学院附属海口医院骨科中心,海南海口570208 [2]海口市骨科与糖尿病医院骨科中心,海南海口570208

出　　处：《临床军医杂志》2023年第9期908-913,共6页Clinical Journal of Medical Officers

基　　金：海南省卫生健康行业课题(19A200123);海南省卫生健康行业课题(20A200518);海南省科技专项资助(ZDYF2020122)。

摘　　要：目的探究长链非编码RNA(LncRNA)HOX转录反义RNA(HOTAIR)通过miR-519c-3p/骨形成蛋白Ⅱ型受体(BMPR2)调控骨关节炎进展的作用及分子机制。方法利用生物信息学网站ENCORI、TargetScanHuman 8.0工具分析预测LncRNA HOTAIR的下游miR-519c-3p以及靶基因BMPR2,并通过双荧光素酶报告及RIP实验验证分子间的相互结合。通过慢病毒转染将miR-519c-3p在细胞中过表达后,将构建好的软骨细胞分为NC组和miR-519c-3p过表达组(miR-519c-3p组)。两组细胞分别应用CCK-8检测、蛋白质印迹、实时荧光定量聚合酶链反应、酶联免疫吸附试验等检测探究LncRNA HOTAIR/miR-519c-3p/BMPR2轴对人软骨细胞骨关节炎的进展的影响。结果与NC组比较,miR-519c-3p组细胞生长变慢,降解酶[降基质金属蛋白酶(MMP)3、MMP9、MMP13、含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶(ADAMTS)4和ADAMTS5]和促炎因子(白细胞介素1β、白细胞介素6和肿瘤坏死因子α)水平显著降低,而COL2A1和SOX9水平升高,差异均有统计学意义(P<0.05)。此外,LncRNA HOTAIR可结合miR-519c-3p,并且BMPR2是miR-519c-3p的下游靶基因,回补实验证实LncRNA HOTAIR可通过miR-519c-3p/BMPR2轴调控人软骨细胞生长、降解酶以及促炎因子的分泌。结论LncRNA HOTAIR可通过miR-519c-3p/BMPR2轴调控骨关节炎的进展。Objective To investigate the role and molecular mechanism of long noncoding RNA(LncRNA)HOX transcript antisense(HOTAIR)in regulating the progression of osteoarthritis through miR-519c-3p/bone morphogenetic protein receptor 2(BMPR2).Methods The ENCORI was used to predict the downstream miR-519c-3p of LncRNA HOTAIR and the TargetScanHuman 8.0 tool was used to predict the target gene BMPR2 of the miR-519c-3p.The intermolecular interactions were verified by dual luciferase reporter and RIP assay.After overexpression of miR-519c-3p by lentiviral transfection,the stably passaged chondrocytes were divided into NC group and miR-519c-3p overexpression group(miR-519c-3p group).The influence of LncRNA HOTAIR/miR-519c-3p/BMPR2 axis on the progression of human chondrocyte osteoarthritis was investigated by CCK-8,western blot,quantitative real-time polymerase chain reaction,enzyme-linked immunosorbent assay.Results Compared with the NC group,the cell growth of miR-519c-3p group slowed down.The levels of degrading enzymes[decreased matrix metalloproteinases(MMP)3,MMP9,MMP13,depolymerized protein-like metalloproteinases(ADAMTS)4 and ADAMTS5 containing platelet-binding protein motifs of typeⅠ]and pro-inflammatory factors(interleukin 1β,interleukin 6 and tumor necrosis factor α)were significantly reduced.The levels of COL2A1 and SOX9 were increased,and the differences were statistically significant(P<0.05).LncRNA HOTAIR could combine with miR-519c-3p,and BMPR2 was a downstream target gene of miR-519c-3p.LncRNA HOTAIR could regulate the growth of human chondrocytes,the secretion of degrading enzymes and pro-inflammatory factors through the miR-519c-3p/BMPR2 axis.Conclusion LncRNA HOTAIR/miR-519c-3p/BMPR2 axis regulates the progression of human chondrocyte osteoarthritis.

关 键 词：骨关节炎 长链非编码RNA HOX转录反义RNA 微小RNA-519c-3p 骨形成蛋白Ⅱ型受体 

分 类 号：R684.3[医药卫生—骨科学]