Department of Joint Surgery, Affiliated Hospital of Jiangsu University, Zhen Jiang, Jiangsu Province, China.
J Gene Med. 2021 Oct;23(10):e3372. doi: 10.1002/jgm.3372. Epub 2021 Jul 16.
Osteoporosis (OP) is a bone disease characterized by reduced amount and quality of bone. This study was designed to explore the role and mechanism of lncRNA IGF2-AS in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).
Human lncRNA and miRNA microarray analyses were performed to measure the differential expression levels of lncRNAs and miRNAs in undifferentiated and osteogenically differentiated BMSCs. lncRNA IGF2-AS, miR-3,126-5p, and KLK4 levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Osteogenic differentiation of BMSCs was assessed by alkaline phosphatase (ALP) staining and Alizarin Red staining (ARS). Protein levels of osterix (Osx), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) were examined by RT-PCR and western blot assays. The binding relationship between miR-3,126-5p and lncRNA IGF2-AS or KLK4 was predicted by TargetScan (http://www.targetscan.org/vert_72/) and then verified with a dual-luciferase reporter assay.
lncRNA IGF2-AS and KLK4 were highly expressed and miR-3,126-5p was weakly expressed in osteogenically differentiated BMSCs. Moreover, lncRNA IGF2-AS overexpression enhanced the osteogenic differentiation of BMSCs. In contrast, lncRNA IGF2-AS knockdown showed the opposite trend. Moreover, miR-3,126-5p overexpression abolished the lncRNA IGF2-AS-mediated osteogenic differentiation of BMSCs. lncRNA IGF2-AS functions as a sponge of miR-3,126-5p to regulate KLK4 expression.
lncRNA IGF2-AS enhances the osteogenic differentiation of BMSCs by modulating the miR-3,126-5p/KLK4 axis, suggesting a promising therapeutic target for bone-related diseases.
骨质疏松症（OP）是一种以骨量和骨质量减少为特征的骨骼疾病。本研究旨在探讨长链非编码 RNA（lncRNA）IGF2-AS 在骨髓间充质干细胞（BMSCs）成骨分化中的作用和机制。
通过人类 lncRNA 和 miRNA 微阵列分析测量未分化和成骨分化的 BMSCs 中 lncRNA 和 miRNA 的差异表达水平。通过实时定量聚合酶链反应（RT-qPCR）测量 lncRNA IGF2-AS、miR-3、126-5p 和 KLK4 的水平。通过碱性磷酸酶（ALP）染色和茜素红染色（ARS）评估 BMSCs 的成骨分化。通过 RT-PCR 和 Western blot 检测osterix（Osx）、骨钙素（OCN）和 runt 相关转录因子 2（RUNX2）的蛋白水平。通过 TargetScan（http://www.targetscan.org/vert_72/）预测 miR-3、126-5p 与 lncRNA IGF2-AS 或 KLK4 的结合关系，并通过双荧光素酶报告基因检测进行验证。
lncRNA IGF2-AS 和 KLK4 在成骨分化的 BMSCs 中高表达，miR-3、126-5p 低表达。此外，lncRNA IGF2-AS 过表达增强了 BMSCs 的成骨分化。相反，lncRNA IGF2-AS 敲低则表现出相反的趋势。此外，miR-3、126-5p 过表达消除了 lncRNA IGF2-AS 介导的 BMSCs 成骨分化。lncRNA IGF2-AS 作为 miR-3、126-5p 的海绵调节 KLK4 的表达。
lncRNA IGF2-AS 通过调节 miR-3、126-5p/KLK4 轴增强 BMSCs 的成骨分化，为骨相关疾病的治疗提供了有前途的靶点。
