Gao Xiaoya, Xue Yun, Yang Kechun
Department of Endocrinology, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu 213161, P.R. China.
Exp Ther Med. 2021 Oct;22(4):1071. doi: 10.3892/etm.2021.10505. Epub 2021 Jul 28.
Accumulating researches indicate that long non-coding RNAs (lncRNAs) participate in human bone mesenchymal stem cells (hBMSCs) osteogenic differentiation. The present study aimed to investigate the underlying molecular mechanisms of long intergenic non-protein coding RNA 899 (LINC00899) in osteoporosis. Therefore, reverse transcription-quantitative PCR was performed to evaluate the expression levels of LINC00899, microRNA (miR)-374a and runt-related transcription factor 2 (RUNX2) in clinical tissues and hBMSCs. The potential interaction between miR-374a and LINC00899 or RUNX2 was predicted utilizing the StarBase software and verified by luciferase reporter and RNA binding protein immunoprecipitation assays. In addition, alkaline phosphatase activity and Alizarin Red S staining were used to evaluate the osteogenic potential of hBMSCs. The results showed that the expression levels of LINC00899 were gradually increased, whilst those of miR-374a were decreased as the osteogenic induction process progresses. Additionally, the expression of LINC00899 was downregulated in the bone tissues of patients with osteoporosis, where LINC00899 knockdown reduced the expression levels of osteogenesis-related genes osteocalcin (OCN), osteopontin (OPN) and RUNX2 in hBMSCs. LINC00899 was also found to directly target miR-374a, thereby inhibiting its expression. Finally, it was predicted that RUNX2 could be directly targeted by miR-374a, such that miR-374a silencing partially abolished the inhibitory effect of LINC00899 knockdown on the expression of RUNX2, OPN and OCN. Overall, findings of the present study suggested that LINC00899 could facilitate the osteogenic differentiation of hBMSCs and prevent osteoporosis by sponging miR-374a to enhance the expression of RUNX2, which provide a potentially useful therapeutic strategy for patients with osteoporosis.
越来越多的研究表明,长链非编码RNA(lncRNAs)参与人类骨髓间充质干细胞(hBMSCs)的成骨分化。本研究旨在探讨长链基因间非编码RNA 899(LINC00899)在骨质疏松症中的潜在分子机制。因此,采用逆转录定量PCR检测临床组织和hBMSCs中LINC00899、微小RNA(miR)-374a和 runt相关转录因子2(RUNX2)的表达水平。利用StarBase软件预测miR-374a与LINC00899或RUNX2之间的潜在相互作用,并通过荧光素酶报告基因和RNA结合蛋白免疫沉淀试验进行验证。此外,采用碱性磷酸酶活性和茜素红S染色评估hBMSCs的成骨潜能。结果显示,随着成骨诱导过程的进展,LINC00899的表达水平逐渐升高,而miR-374a的表达水平则降低。此外,骨质疏松症患者骨组织中LINC00899的表达下调,敲低LINC00899可降低hBMSCs中成骨相关基因骨钙素(OCN)、骨桥蛋白(OPN)和RUNX2的表达水平。还发现LINC00899直接靶向miR-374a,从而抑制其表达。最后,预测RUNX2可能是miR-374a的直接靶点,因此沉默miR-374a可部分消除敲低LINC00899对RUNX2、OPN和OCN表达的抑制作用。总体而言,本研究结果表明,LINC00899可通过吸附miR-374a以增强RUNX2的表达,从而促进hBMSCs的成骨分化并预防骨质疏松症,这为骨质疏松症患者提供了一种潜在的有效治疗策略。