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miR-1827 inhibits osteogenic differentiation by targeting IGF1 in MSMSCs

View Article: PubMed Central - PubMed

ABSTRACT

We recently reported that maxillary sinus membrane stem cells (MSMSCs) have osteogenic potential. However, the biological mechanisms of bone formation remain unclear. In this study, we investigated the role and mechanisms of microRNAs (miRNAs) in the osteogenic differentiation of MSMSCs. The expression of miRNAs was determined in differentiated MSMSCs by comprehensive miRNA microarray analysis and quantitative RT-PCR (qRT-PCR). We selected miR-1827 for functional follow-up studies to explore its significance in MSMSCs. Here, miR-1827 was found to be up-regulated during osteogenic differentiation of MSMSCs. Over expression of miR-1827 inhibited osteogenic differentiation of MSMSCs in vitro, whereas the repression of miR-1827 greatly promoted cell differentiation. Further experiments confirmed that insulin-like growth factor 1 (IGF1) is a direct target of miR-1827. miR-1827 inhibited osteogenic differentiation partially via IGF1, which in turn is a positive regulator of osteogenic differentiation. Moreover, miR-1827 suppressed ectopic bone formation and silencing of miR-1827 led to increased bone formation in vivo. In summary, this study is the first to demonstrate that miR-1827 can regulate osteogenic differentiation. The increase in miR-1827 expression observed during osteogenesis is likely a negative feedback mechanism, thus offering a potential therapeutic target to address inadequate bone volume for dental implantation through inhibiting miR-1827.

No MeSH data available.


IGF1 is a target of miR-1827 in MSMSCs and BMSSCs.qRT-PCR analysis of candidate gene (IGF1 and Grb2) expression in MSMSCs after transfection with the miR-1827 mimic or negative control for 48 h. (b) A schematic of the luciferase reporter containing the putative binding sites for miR-1827 in the IGF1 3′-UTR. The sequences of miR-1827, its putative binding sites, and their respective IGF1 3′-UTR mutants are presented. The positions of putative binding sites are labeled. (c,d) Dual luciferase activity in transfected MSMSCs and BMSSCs. (e) Western blot analysis of IGF1 protein levels in MSMSCs after treatment with miR-1827 mimic, miR-1827 inhibitor or their respective negative controls for 48 h. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.
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f3: IGF1 is a target of miR-1827 in MSMSCs and BMSSCs.qRT-PCR analysis of candidate gene (IGF1 and Grb2) expression in MSMSCs after transfection with the miR-1827 mimic or negative control for 48 h. (b) A schematic of the luciferase reporter containing the putative binding sites for miR-1827 in the IGF1 3′-UTR. The sequences of miR-1827, its putative binding sites, and their respective IGF1 3′-UTR mutants are presented. The positions of putative binding sites are labeled. (c,d) Dual luciferase activity in transfected MSMSCs and BMSSCs. (e) Western blot analysis of IGF1 protein levels in MSMSCs after treatment with miR-1827 mimic, miR-1827 inhibitor or their respective negative controls for 48 h. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.

Mentions: To further explore the mechanism by which miR-1827 regulates osteogenic differentiation, we examined the predicted targets of miR-1827 using TargetScan, miRanda and miRDB software. Based on these analyses, several potential targets meeting this criterion were identified (Supplementary Figure 1). From these genes, we selected insulin-like growth factor 1 (IGF1) and growth factor receptor binding protein 2 (Grb2) for further study, because these proteins are potentially involved in osteogenic differentiation through MAPK signaling pathways1920. Therefore, we examined the mRNA expression of each target in response to the miR-1827 mimic and miR-1827 inhibitor. We observed that the mRNA expression of IGF1, but not Grb2, was significantly changed (Fig. 3a). Therefore, IGF1 was selected as the better candidate between the two genes.


miR-1827 inhibits osteogenic differentiation by targeting IGF1 in MSMSCs
IGF1 is a target of miR-1827 in MSMSCs and BMSSCs.qRT-PCR analysis of candidate gene (IGF1 and Grb2) expression in MSMSCs after transfection with the miR-1827 mimic or negative control for 48 h. (b) A schematic of the luciferase reporter containing the putative binding sites for miR-1827 in the IGF1 3′-UTR. The sequences of miR-1827, its putative binding sites, and their respective IGF1 3′-UTR mutants are presented. The positions of putative binding sites are labeled. (c,d) Dual luciferase activity in transfected MSMSCs and BMSSCs. (e) Western blot analysis of IGF1 protein levels in MSMSCs after treatment with miR-1827 mimic, miR-1827 inhibitor or their respective negative controls for 48 h. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.
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Related In: Results  -  Collection

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f3: IGF1 is a target of miR-1827 in MSMSCs and BMSSCs.qRT-PCR analysis of candidate gene (IGF1 and Grb2) expression in MSMSCs after transfection with the miR-1827 mimic or negative control for 48 h. (b) A schematic of the luciferase reporter containing the putative binding sites for miR-1827 in the IGF1 3′-UTR. The sequences of miR-1827, its putative binding sites, and their respective IGF1 3′-UTR mutants are presented. The positions of putative binding sites are labeled. (c,d) Dual luciferase activity in transfected MSMSCs and BMSSCs. (e) Western blot analysis of IGF1 protein levels in MSMSCs after treatment with miR-1827 mimic, miR-1827 inhibitor or their respective negative controls for 48 h. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.
Mentions: To further explore the mechanism by which miR-1827 regulates osteogenic differentiation, we examined the predicted targets of miR-1827 using TargetScan, miRanda and miRDB software. Based on these analyses, several potential targets meeting this criterion were identified (Supplementary Figure 1). From these genes, we selected insulin-like growth factor 1 (IGF1) and growth factor receptor binding protein 2 (Grb2) for further study, because these proteins are potentially involved in osteogenic differentiation through MAPK signaling pathways1920. Therefore, we examined the mRNA expression of each target in response to the miR-1827 mimic and miR-1827 inhibitor. We observed that the mRNA expression of IGF1, but not Grb2, was significantly changed (Fig. 3a). Therefore, IGF1 was selected as the better candidate between the two genes.

View Article: PubMed Central - PubMed

ABSTRACT

We recently reported that maxillary sinus membrane stem cells (MSMSCs) have osteogenic potential. However, the biological mechanisms of bone formation remain unclear. In this study, we investigated the role and mechanisms of microRNAs (miRNAs) in the osteogenic differentiation of MSMSCs. The expression of miRNAs was determined in differentiated MSMSCs by comprehensive miRNA microarray analysis and quantitative RT-PCR (qRT-PCR). We selected miR-1827 for functional follow-up studies to explore its significance in MSMSCs. Here, miR-1827 was found to be up-regulated during osteogenic differentiation of MSMSCs. Over expression of miR-1827 inhibited osteogenic differentiation of MSMSCs in vitro, whereas the repression of miR-1827 greatly promoted cell differentiation. Further experiments confirmed that insulin-like growth factor 1 (IGF1) is a direct target of miR-1827. miR-1827 inhibited osteogenic differentiation partially via IGF1, which in turn is a positive regulator of osteogenic differentiation. Moreover, miR-1827 suppressed ectopic bone formation and silencing of miR-1827 led to increased bone formation in vivo. In summary, this study is the first to demonstrate that miR-1827 can regulate osteogenic differentiation. The increase in miR-1827 expression observed during osteogenesis is likely a negative feedback mechanism, thus offering a potential therapeutic target to address inadequate bone volume for dental implantation through inhibiting miR-1827.

No MeSH data available.