Abstract
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/β-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.
Highlights
Many studies have shown that long noncoding RNAs are closely related to the stimulation of osteogenic differentiation of adipose-derived stem cells (ADSCs) and the prevention of osteoporosis
ADSCs could successfully differentiate into osteoblasts (ARS, Fig. 1b), Fig. 2 The volcano plots of the significantly differentially expressed (P < 0.05, fold change > 2) long noncoding RNAs (lncRNAs) (a)
LncRNA LINC00314 is upregulated during ADSCs osteogenic differentiation To further determine which lncRNA and microRNA participant into the process of ADSCs osteogenic differentiation, differentially altered lncRNA and microRNA in non-induced and induced ADSCs were analyzed further (Fig. 2a)
Summary
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. Osteoporosis is a serious degenerative disease characterized by the reduction of bone mass and destruction of the bone microstructure [1, 2]. Wu et al [10] analyzed the differentially expressed long noncoding RNAs in the normal and induced groups and found that HIF1A-AS2 has a positive role in promoting ADSCs osteogenic differentiation through the PI3K/Akt signaling pathway. No study has explored the role and mechanism of LINC00314 in the regulation of ADSC osteogenic differentiation
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