Long noncoding RNA LINC00314 facilitates osteogenic differentiation of adipose-derived stem cells through the hsa-miR-129-5p/GRM5 axis via the Wnt signaling pathway
Long noncoding RNA LINC00314 facilitates osteogenic differentiation of adipose-derived stem cells through the hsa-miR-129-5p/GRM5 axis via the Wnt signaling pathway
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DOI：
10.1186/s13287-020-01754-z
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发表时间：
2020-06-17
影响因子：
7.5
通讯作者：
Feng, Guo-jun
中科院分区：
文献类型：
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作者：
Shi, Zheng-liang;Zhang, Hua;Feng, Guo-jun
背景许多研究表明，长非编码RNA(Long Non Coding RNAs，LncRNAs)与促进脂肪干细胞成骨分化、预防骨质疏松密切相关。本研究旨在研究新的LncRNA，探讨LINC00314/miR-129-5p/GRM5轴在调控ADSC成骨分化中的作用及其分子机制。方法对正常和成骨诱导的ADSC进行LncRNA和miRNA测序。用R软件分析LINC00314、miR-129-5p和GRM5在成骨诱导过程中的表达，用RT-PCR方法检测成骨诱导过程中LINC00314、miR-129-5p和GRM5的相对表达。将pcDNA3.1-sh-LINC00314和agomiR-129-5p分别导入ADSCs。用茜素红(ARS)染色和碱性磷酸酶(ALP)染色鉴定LINC00314/miR-129-5p/GRM5轴调控ADSC成骨分化的机制。LINC00314和GRM5模拟物增加成骨分化的早期和晚期标志物，不仅表现为ALP活性显著升高，还表现为钙沉积增加，而miR-129-5P模拟物的作用则相反。LINC00314通过荧光素酶报告基因直接靶向miR-129-5p，miR-129-5p抑制GRM5的表达。结论LINC00314/miR-129-5p/GRM5在ADSCs的成骨分化中起重要作用，LINC00314/miR-129-5p/GRM5可能是成骨相关疾病的新机制。
BackgroundMany 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.MethodsLncRNA 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.ResultsLINC00314 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/beta -catenin signaling pathway.ConclusionsLINC00314 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.