Orthopedic Department, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Eur Rev Med Pharmacol Sci. 2019 Dec;23(23):10195-10203. doi: 10.26355/eurrev_201912_19654.
To investigate the expression of long noncoding RNA (LncRNA) MIRG and its potential functions in regulating osteoclastogenesis and bone resorption function through modulating miR-1897 in bone marrow macrophages (BMMs).
qRT-PCR was performed to detect the expressions of MIRG and its co-expression mRNA NFATc1 at different stages during osteoclastogenesis. The CCK-8 assay was performed to evaluate cell proliferation and differentiation. The correlation between miR-1897 and MIRG was detected by statistical analysis. Bioinformatics and luciferase assay were performed to explore the interaction and binding site of MIRG and miR-1897. We also cloned the mice NFATc1 3'-UTR into the luciferase reporter vector and constructed miR-1897 binding mutants to validate the inhibited regulation of miR-1897 to the expression of NFATc1.
Results showed that expressions of MIRG and NFATc1 were upregulated during osteoclastogenesis. qRT-PCR and CCK-8 assay showed that MIRG expression is associated with osteoclastogenesis and bone resorption. The bioinformatics prediction and luciferase assay suggested that by interacting with miR-1897, MIRG acts as a molecular sponge for the miR-1897 target NFATc1, to partly modulate the inhibitory effect of miR-1897 on NFATc1.
We found that lncRNA-MIRG was upregulated in osteoclasts, which could promote osteoclastogenesis and bone resorption function as a molecular sponge by modulating the inhibitory effect of miR-1897 on NFATc1.
通过调节骨髓巨噬细胞(BMM)中的 miR-1897,研究长链非编码 RNA(LncRNA)MIRG 的表达及其在调节破骨细胞生成和骨吸收功能中的潜在作用。
qRT-PCR 检测破骨细胞生成过程中不同阶段 MIRG 及其共表达 mRNA NFATc1 的表达。CCK-8 法检测细胞增殖和分化。统计分析检测 miR-1897 与 MIRG 的相关性。生物信息学和荧光素酶报告基因实验检测 MIRG 和 miR-1897 的相互作用和结合位点。我们还将小鼠 NFATc1 3'-UTR 克隆到荧光素酶报告基因载体中,并构建 miR-1897 结合突变体,以验证 miR-1897 对 NFATc1 表达的抑制调节作用。
结果表明,MIRG 和 NFATc1 的表达在破骨细胞生成过程中上调。qRT-PCR 和 CCK-8 检测结果表明,MIRG 的表达与破骨细胞生成和骨吸收有关。生物信息学预测和荧光素酶报告基因实验表明,通过与 miR-1897 相互作用,MIRG 作为 miR-1897 靶 NFATc1 的分子海绵,部分调节 miR-1897 对 NFATc1 的抑制作用。
我们发现,lncRNA-MIRG 在破骨细胞中上调,可通过调节 miR-1897 对 NFATc1 的抑制作用,作为分子海绵促进破骨细胞生成和骨吸收功能。