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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.


Inhibition of osteogenic differentiation by miR-1827 partially depends on IGF1.(a,b) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (c) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (d,e) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (f,g) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (h) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (i,j) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.
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f5: Inhibition of osteogenic differentiation by miR-1827 partially depends on IGF1.(a,b) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (c) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (d,e) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (f,g) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (h) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (i,j) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.

Mentions: To further confirm that the inhibition of osteogenic differentiation by miR-1827 depends on IGF1, we co-transfected MSMSCs with the miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. The co-transfection of miR-1827 inhibitor with siRNA-IGF1 partially attenuated the miR-1827 inhibitor-induced increase in Runx2 and OPN mRNA expression as indicated by qRT-PCR (Fig. 5a,b). The increase in ALP activity induced by miR-1827 inhibitor was also partially inhibited (Fig. 5c). These changes persisted at the protein level, where co-transfection of miR-1827 inhibitor with siRNA-IGF1 partially blocked the increase in Runx2 and OPN protein expression induced by the miR-1827 inhibitor (Fig. 5d,e). Moreover, we transfected miR-1827 mimic into cells infected with ADIGF1 or ADGFP. The co-transfection of miR-1827 mimic with ADIGF1 partially blocked the miR-1827 mimic-induced reduction of Runx2 and OPN mRNA expression (Fig. 5f,g). The reduction of ALP activity induced by the miR-1827 inhibitor was also partially blocked (Fig. 5h). Similar alterations in Runx2 and OPN expression were also observed at the protein level (Fig. 5i,j).


miR-1827 inhibits osteogenic differentiation by targeting IGF1 in MSMSCs
Inhibition of osteogenic differentiation by miR-1827 partially depends on IGF1.(a,b) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (c) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (d,e) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (f,g) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (h) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (i,j) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.
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f5: Inhibition of osteogenic differentiation by miR-1827 partially depends on IGF1.(a,b) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (c) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (d,e) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. (f,g) qRT-PCR analysis of Runx2 and OPN mRNA expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (h) Analysis of ALP activity in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. (i,j) Western blot analysis of Runx2 and OPN protein expression in MSMSCs after co-transfection with miR-1827 mimic and ADIGF1 or their respective negative controls. For each group, values are the mean ± SD; n = 3, *P < 0.05, **P < 0.01. NS, not significant.
Mentions: To further confirm that the inhibition of osteogenic differentiation by miR-1827 depends on IGF1, we co-transfected MSMSCs with the miR-1827 inhibitor and siRNA-IGF1 or their respective negative controls. The co-transfection of miR-1827 inhibitor with siRNA-IGF1 partially attenuated the miR-1827 inhibitor-induced increase in Runx2 and OPN mRNA expression as indicated by qRT-PCR (Fig. 5a,b). The increase in ALP activity induced by miR-1827 inhibitor was also partially inhibited (Fig. 5c). These changes persisted at the protein level, where co-transfection of miR-1827 inhibitor with siRNA-IGF1 partially blocked the increase in Runx2 and OPN protein expression induced by the miR-1827 inhibitor (Fig. 5d,e). Moreover, we transfected miR-1827 mimic into cells infected with ADIGF1 or ADGFP. The co-transfection of miR-1827 mimic with ADIGF1 partially blocked the miR-1827 mimic-induced reduction of Runx2 and OPN mRNA expression (Fig. 5f,g). The reduction of ALP activity induced by the miR-1827 inhibitor was also partially blocked (Fig. 5h). Similar alterations in Runx2 and OPN expression were also observed at the protein level (Fig. 5i,j).

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.