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环状 RNA_009934 通过沉默 miR-5107 诱导破骨细胞骨吸收。

CircRNA_009934 induces osteoclast bone resorption via silencing miR-5107.

机构信息

Department of Endocrinology, Shanxi Provincial People's Hospital, Taiyuan City, Shanxi Province, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Jul;24(14):7580-7588. doi: 10.26355/eurrev_202007_22256.

DOI:10.26355/eurrev_202007_22256
PMID:32744684
Abstract

OBJECTIVE

We aimed to explore the expression of circRNA_009934 in osteoclast, as well as its potential roles in regulating osteoclastogenesis and bone resorption via regulating miR-5107.

MATERIALS AND METHODS

We performed qRT-PCR analysis to examine the expression of circRNA_009934 in osteoclast in distinctive stages. We used CCK-8 assay to detect the cell proliferation ability. Correlation analysis between the expression levels of circRNA_009934 and miR-5107 was performed using statistical analysis. Bioinformatics prediction was performed to predict the binding site of circRNA_009934 and miR-5107, subsequently followed by Luciferase assay for validation. The mice TRAF6 3'-UTR were cloned into the Luciferase reporter vector and miR-5107 binding mutants were constructed to validate the inhibited regulation of miR-5107 to the expression of TRAF6.

RESULTS

Our results showed that expression of circRNA_009934 was increased during osteoclast differentiation. CircRNA_009934 expression was closely correlated with osteoclastogenesis and bone resorption activity. Bioinformatics prediction and Luciferase assay demonstrated that circRNA_009934 served as a ceRNA of miR-5107 and regulated its downstream TRAF6 expression.

CONCLUSIONS

We first demonstrated that circRNA_009934 expression was increased in osteoclasts, which promoted osteoclastogenesis by serving as a ceRNA of miR-5107 and regulated the expression of TRAF6.

摘要

目的

我们旨在探索 circRNA_009934 在破骨细胞中的表达,以及通过调节 miR-5107 来调节破骨细胞生成和骨吸收的潜在作用。

材料和方法

我们通过 qRT-PCR 分析来检测不同阶段破骨细胞中 circRNA_009934 的表达。我们使用 CCK-8 检测细胞增殖能力。通过统计分析进行 circRNA_009934 和 miR-5107 表达水平之间的相关性分析。进行生物信息学预测以预测 circRNA_009934 和 miR-5107 的结合位点,随后进行 Luciferase 测定进行验证。将小鼠 TRAF6 3'-UTR 克隆到 Luciferase 报告载体中,并构建 miR-5107 结合突变体以验证 miR-5107 对 TRAF6 表达的抑制调节作用。

结果

我们的结果表明,circRNA_009934 的表达在破骨细胞分化过程中增加。circRNA_009934 的表达与破骨细胞生成和骨吸收活性密切相关。生物信息学预测和 Luciferase 测定表明,circRNA_009934 作为 miR-5107 的 ceRNA 并调节其下游 TRAF6 表达。

结论

我们首次证明 circRNA_009934 在破骨细胞中表达增加,通过作为 miR-5107 的 ceRNA 促进破骨细胞生成,并调节 TRAF6 的表达。

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