Many studies have shown that long noncoding RNAs (lncRNAs) are closely related to the stimulation of osteogenic differentiation of adipose-derived stem cells (ADSCs) and the prevention of osteoporosis. Current research aimed to investigate the novel lncRNA and explored the function and molecular mechanism of the LINC00314/miR-129-5p/GRM5 axis in regulating osteogenic differentiation of ADSCs. LncRNA and miRNA sequencing was performed in normal and osteogenic differentiation-induced ADSCs (osteogenic group). Abnormally expressed lncRNAs and miRNAs were obtained by the R software and the relative expression of LINC00314, miR-129-5p, and GRM5 during osteogenic induction was measured by RT-PCR. ADSCs were then transfected with pcDNA3.1-sh-LINC00314 and agomiR-129-5p. Alizarin red staining (ARS) and alkaline phosphatase (ALP) staining were performed to identify the mechanism of the LINC00314/miR-129-5p/GRM5 axis in regulating osteogenic differentiation of ADSCs. LINC00314 was significantly upregulated in the group of osteogenic-induced ADSCs. LINC00314 and GRM5 mimics increased the early and late markers of osteogenic differentiation, which manifest in not only the markedly increased ALP activity but also higher calcium deposition, while miR-129-5p mimic had the opposite effects. LINC00314 directly targeted miR-129-5p through luciferase reporter assay, and miR-129-5p suppressed GRM5 expression. Moreover, the LINC00314/miR-129-5p/GRM5 regulatory axis activated the Wnt/β-catenin signaling pathway. LINC00314 confers contributory function in the osteogenic differentiation of ADSCs and thus the LINC00314/miR-129-5p/GRM5 axis may be a novel mechanism for osteogenic-related disease.

中文翻译：

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长链非编码 RNA LINC00314 通过 Wnt 信号通路通过 hsa-miR-129-5p/GRM5 轴促进脂肪干细胞的成骨分化。

许多研究表明，长链非编码RNA（lncRNAs）与刺激脂肪干细胞（ADSCs）的成骨分化和预防骨质疏松症密切相关。目前的研究旨在研究新型lncRNA，并探索LINC00314/miR-129-5p/GRM5轴在调节ADSCs成骨分化中的功能和分子机制。在正常和成骨分化诱导的 ADSC（成骨组）中进行 LncRNA 和 miRNA 测序。通过R软件获得异常表达的lncRNA和miRNA，并通过RT-PCR测量LINC00314、miR-129-5p和GRM5在成骨诱导过程中的相对表达。然后用 pcDNA3.1-sh-LINC00314 和 agomiR-129-5p 转染 ADSC。进行茜素红染色 (ARS) 和碱性磷酸酶 (ALP) 染色以确定 LINC00314/miR-129-5p/GRM5 轴在调节 ADSCs 成骨分化中的机制。LINC00314 在成骨诱导的 ADSC 组中显着上调。LINC00314 和 GRM5 模拟物增加了成骨分化的早期和晚期标志物，这不仅表现在 ALP 活性显着增加，而且钙沉积更高，而 miR-129-5p 模拟物具有相反的效果。LINC00314 通过荧光素酶报告基因检测直接靶向 miR-129-5p，miR-129-5p 抑制 GRM5 表达。此外，LINC00314/miR-129-5p/GRM5 调节轴激活了 Wnt/β-catenin 信号通路。