长非编码RNA SNAI3-AS1调控人软骨细胞C28/I2增殖能力及退变的机制研究  

Mechanism of Long Non-coding RNA SNAI3-AS1 on Proliferation and Degeneration of Human Chondrocytes

作　　者：王婵 王晶 何学智 王璟 苗登顺[1] WANG Chan;WANG Jing;HE Xue-zhi;WANG Jing;MIAO Deng-shun(Department of Anatomy,Histology and Embryology,Nanjing Medical University,Nanjing,Jiangsu,211100,China)

机构地区：[1]南京医科大学基础医学院人体解剖与组织胚胎学系骨与干细胞研究中心,江苏南京211100

出　　处：《现代生物医学进展》2023年第14期2601-2608,共8页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(8217091038);国家青年科学基金项目(8210115154)。

摘　　要：目的:为了探究长非编码RNA SNAI3-AS1(LncRNA SNAI3-AS1,即SNAI3-AS1)在骨性关节炎(osteoarthritis,OA)进展中的作用与机制。方法:通过全转录组测序筛选出在OA中差异表达的lncRNA SNAI3-AS1,并通过实时荧光定量PCR(qRT-PCR)检测SNAI3-AS1在软骨细胞退变模型中的表达情况。在软骨细胞C28/I2中分别转染SNAI3-AS1特异性si RNA或真核过表达质粒,分别敲低或过表达SNAI3-AS1,通过MTT、平板克隆形成和EdU掺入实验检测细胞增殖活力,Western Blot检测炎症和细胞外基质蛋白的表达情况。通过生物信息学网站预测SNAI3-AS1相互作用的miRNA和下游靶基因,并通过双荧光素酶报告基因和RIP实验进行验证。结果:相较于正常软骨细胞,SNAI3-AS1的表达水平在OA中显著下调。敲低正常软骨细胞中SNAI3-AS1的表达后,软骨细胞的增殖能力减弱并促进了软骨细胞的退变,而在OA模型的软骨细胞中过表达SNAI3-AS1后,软骨细胞的增殖活力加强并抑制了软骨细胞的退变。在机制上,SNAI3-AS1可充当竞争性内源性RNA(ceRNA),经海绵吸附miR-2278间接上调PRELP,发挥促进软骨细胞增殖和抑制其退变的作用。结论:LncRNA SNAI3-AS1通过LncRNA SNAI3-AS1/miR-2278/PRELP轴参与骨性关节炎的发生发展过程。Objective:In order to explore the function and influence of long non-coding RNA SNAI3-AS1(LncRNA SNAI3-AS1,SNAI3-AS1)in the progression of osteoarthritis(OA).Methods:LncRNA SNAI3-AS1 was screened by whole-transcriptome sequencing,and its expression in cell model of chondrocyte degeneration was detected by real-time quantitative fluorescence polymerase chain reaction(qRT-PCR).Transfecting SNAI3-AS1-specific si-RNA or eukaryotic overexpression plasmid in chondrocytes to knock down or overexpress SNAI3-AS1 separately.The proliferation activity of chondrocytes was detected by MTT assay,plate colony assay and EdU assay,and the expression of inflammatory and extracellular matrix proteins was detected by Western Blot.The lncRNA-miRNA and miRNA-mRNA interactions were predicted by bioinformatics website,and verified by dual luciferase reporter gene and RIP assay.Results:Compared with normal human chondrocytes,the expression level of SNAI3-AS1 was significantly down-regulated in OA.And knockdown of SNAI3-AS1 in chondrocytes can reduce chondrocyte proliferation and promote chondrocyte degeneration,while overexpression of SNAI3-AS1 in chondrocytes of OA model can enhance chondrocyte proliferation and inhibit chondrocyte degeneration.In terms of mechanism,SNAI3-AS1 can act as a competitive endogenous RNA(ceRNA),which indirectly up-regulates PRELP by sponging miR-2278,and plays a role in promoting chondrocyte proliferation and inhibiting its degeneration.Conclusion:LncRNA SNAI3-AS1 is involved in the occurrence and development of osteoarthritis through the lncRNA SNAI3-AS1/miR-2278/PRELP axis.

关 键 词：LncRNA SNAI3-AS1 骨性关节炎 细胞增殖 细胞退变 

分 类 号：R-33[医药卫生] R684.3