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长链非编码 RNA XIXT 通过靶向 miRNA-30a-5p 促进骨髓间充质干细胞的成骨分化并缓解骨质疏松症的进展。

LncRNA XIXT promotes osteogenic differentiation of bone mesenchymal stem cells and alleviates osteoporosis progression by targeting miRNA-30a-5p.

机构信息

Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Oct;23(20):8721-8729. doi: 10.26355/eurrev_201910_19266.

Abstract

OBJECTIVE

To uncover the role of XIXT in influencing the osteogenesis of hBMSCs by adsorbing microRNA-30a-5p (miRNA-30a-5p) to upregulate RUNX2.

PATIENTS AND METHODS

The serum samples were collected from osteoporosis patients and normal people. hBMSCs were isolated from femoral head tissues. The serum levels of XIXT and miRNA-30a-5p were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The expression levels and activities of the osteogenic differentiation-related genes in hBMSCs after transfection of sh-XIXT, sh-RUNX2, miRNA-30a-5p mimic, and inhibitor were detected by qRT-PCR, Western blot, ALP activity assay, and alizarin red staining. The Dual-Luciferase Reporter Gene Assay was performed to confirm the binding of XIXT to miRNA-30a-5p, as well as the binding of miRNA-30a-5p to RUNX2.

RESULTS

LncRNA XIXT was significantly downregulated, and miRNA-30a-5p was upregulated in the serum of osteoporosis patients. The osteogenic differentiation-related genes (ALP, RUNX2) and XIXT were markedly upregulated in a time-dependent manner, while miRNA-30a-5p level gradually decreased in hBMSCs with the prolongation of osteogenesis. The knockdown of XIXT inhibited the osteogenic differentiation of hBMSCs. In hBMSCs, XIXT regulated RUNX2 expression by targeting miRNA-30a-5p. The knockdown of miRNA-30a-5p partially reversed the inhibitory effect of XIXT on the osteogenesis of hBMSCs. However, the downregulated RUNX2 reversed the promotive effect of miRNA-30a-5p on the osteogenesis of hBMSCs.

CONCLUSIONS

LncRNA XIXT upregulated RUNX2 by absorbing miRNA-30a-5p, and thus induced hBMSCs osteogenesis to alleviate osteoporosis.

摘要

目的

通过吸附 microRNA-30a-5p(miRNA-30a-5p)来上调 RUNX2,揭示 XIXT 在影响 hBMSCs 成骨中的作用。

方法

收集骨质疏松症患者和正常人的血清样本。从股骨头组织中分离 hBMSCs。通过实时定量聚合酶链反应(qRT-PCR)测定 hBMSCs 转染 sh-XIXT、sh-RUNX2、miRNA-30a-5p 模拟物和抑制剂后血清中 XIXT 和 miRNA-30a-5p 的水平。通过 qRT-PCR、Western blot、碱性磷酸酶(ALP)活性测定和茜素红染色检测 hBMSCs 成骨分化相关基因的表达水平和活性。双荧光素酶报告基因检测验证 XIXT 与 miRNA-30a-5p 的结合以及 miRNA-30a-5p 与 RUNX2 的结合。

结果

骨质疏松症患者血清中 lncRNA XIXT 明显下调,miRNA-30a-5p 上调。成骨分化相关基因(ALP、RUNX2)和 XIXT 的表达水平随时间呈明显上升趋势,而 hBMSCs 中 miRNA-30a-5p 水平随成骨时间延长逐渐下降。XIXT 下调抑制 hBMSCs 的成骨分化。在 hBMSCs 中,XIXT 通过靶向 miRNA-30a-5p 调节 RUNX2 表达。miRNA-30a-5p 下调部分逆转了 XIXT 对 hBMSCs 成骨的抑制作用。然而,下调的 RUNX2 逆转了 miRNA-30a-5p 对 hBMSCs 成骨的促进作用。

结论

lncRNA XIXT 通过吸附 miRNA-30a-5p 上调 RUNX2,从而诱导 hBMSCs 成骨,缓解骨质疏松症。

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