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FGD5-AS1 通过靶向 miR-506-3p/BMP7 轴促进人骨髓间充质干细胞的成骨分化。

FGD5-AS1 facilitates the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells via targeting the miR-506-3p/BMP7 axis.

机构信息

Department of Spinal Surgery, Changzhou Hospital of Traditional Chinese Medicine, No. 25 Heping North Road, Changzhou, Jiangsu, 213000, P.R. China.

出版信息

J Orthop Surg Res. 2021 Nov 12;16(1):665. doi: 10.1186/s13018-021-02694-x.

DOI:10.1186/s13018-021-02694-x
PMID:34772438
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8588622/
Abstract

BACKGROUND

Osteoporosis is a systemic disease characterized by impaired bone formation, increased bone resorption, and brittle bone fractures. The osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is considered to be a vital process for bone formation. Numerous studies have reported that long non-coding RNAs (lncRNAs) are involved in the osteogenic differentiation of hBMSCs. The present study aimed to investigate the effect of FGD5 antisense RNA 1 (FGD5-AS1) on osteogenic differentiation.

METHODS

RT-qPCR was performed to detect the expression of FGD5-AS1, miR-506-3p, and osteogenesis-related genes OCN, OPN, OSX, and RUNX2. Western blotting was carried out to detect the protein levels of osteogenesis-related markers. In addition, the regulatory effect of FGD5-AS1 on osteogenic differentiation was detected through alkaline phosphatase (ALP) activity, Alizarin Red S (ARS) staining, and Cell Counting Kit-8 (CCK-8). Bioinformatics analysis and luciferase reporter assay were used to predict and validate the interaction between FGD5-AS1 and miR-506-3p as well as miR-506-3p and bone morphogenetic protein 7 (BMP7).

RESULTS

The RT-qPCR analysis revealed that FGD5-AS1 was upregulated in hBMSCs following induction of osteogenic differentiation. In addition, FGD5-AS1 knockdown attenuated hBMSC viability and osteogenic differentiation. Bioinformatics analysis and luciferase reporter assays verified that FGD5-AS1 could directly interact with microRNA (miR)-506-3p. Furthermore, miR-506-3p could directly target the 3'-untranslated region (3'-UTR) of BMP7. Additionally, functional assays demonstrated that miR-506-3p silencing could restore the suppressive effect of FGD5-AS1 knockdown on osteogenic differentiation and viability of hBMSCs, and miR-506-3p could attenuate osteogenic differentiation via targeting BMP7.

CONCLUSIONS

Taken together, the results of the present study suggested that FGD5-AS1 could positively regulate the osteogenic differentiation of hBMSCs via targeting the miR-506-3p/BMP7 axis.

摘要

背景

骨质疏松症是一种以骨形成受损、骨吸收增加和脆性骨折为特征的系统性疾病。人骨髓间充质干细胞（hBMSCs）的成骨分化被认为是骨形成的重要过程。许多研究报道长链非编码 RNA（lncRNA）参与 hBMSCs 的成骨分化。本研究旨在探讨 FGD5 反义 RNA 1（FGD5-AS1）对成骨分化的影响。

方法

采用 RT-qPCR 检测 FGD5-AS1、miR-506-3p 及成骨相关基因 OCN、OPN、OSX、RUNX2 的表达。采用 Western blot 检测成骨相关标志物的蛋白水平。此外，通过碱性磷酸酶（ALP）活性、茜素红 S（ARS）染色和细胞计数试剂盒-8（CCK-8）检测 FGD5-AS1 对成骨分化的调节作用。通过生物信息学分析和荧光素酶报告基因实验预测和验证 FGD5-AS1 与 miR-506-3p 以及 miR-506-3p 与骨形态发生蛋白 7（BMP7）之间的相互作用。

结果

RT-qPCR 分析显示，在 hBMSCs 诱导成骨分化后，FGD5-AS1 上调。此外，FGD5-AS1 敲低降低了 hBMSC 的活力和成骨分化。生物信息学分析和荧光素酶报告基因实验验证了 FGD5-AS1 可以直接与 microRNA（miR）-506-3p 相互作用。此外，miR-506-3p 可以直接靶向 BMP7 的 3'-非翻译区（3'-UTR）。此外，功能分析表明，miR-506-3p 沉默可以恢复 FGD5-AS1 敲低对 hBMSCs 成骨分化和活力的抑制作用，miR-506-3p 可以通过靶向 BMP7 来抑制成骨分化。

结论

综上所述，本研究结果表明，FGD5-AS1 可以通过靶向 miR-506-3p/BMP7 轴正向调节 hBMSCs 的成骨分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/3ccf38ec54e6/13018_2021_2694_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/dca6f1dc6c68/13018_2021_2694_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/099b69153324/13018_2021_2694_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/1ef6cbbaf69f/13018_2021_2694_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/e549ecc51272/13018_2021_2694_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/564ec57092db/13018_2021_2694_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/3ccf38ec54e6/13018_2021_2694_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/dca6f1dc6c68/13018_2021_2694_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/099b69153324/13018_2021_2694_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/1ef6cbbaf69f/13018_2021_2694_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/e549ecc51272/13018_2021_2694_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/564ec57092db/13018_2021_2694_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c517/8588622/3ccf38ec54e6/13018_2021_2694_Fig6_HTML.jpg

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