BMP9-induced Osteogenic Differentiation is Partially Inhibited by miR-30a in Mesenchymal Stem Cell Line C3H10 T1/2

Objective:To study the role of miR-30a in bone morphogenetic protein 9(BMP9) induced osteogenic differentiation of mesenchymal stem cell line C3H10T1 /2.Method:C3H10T1 /2 cells were treated with Adenovirus GFP and BMP9,the early osteogenic marker Alkaline phosphatase(ALP) activity was detected by ALP quantitative assay and staining assay,the later osteogenic marker calcium precipitation was detected by Alizarin Red S staining,OPN was another later marker,which was detected by PCR and Western blot.The expression of miR-30a in C3H10T1 /2 cells quantified by Real-time PCR.Effects of miR-30a on BMP9-induced osteogenic differentiation of C3H10T1 /2 cells was detected by ALP,OPN and calcium precipitation.Runx2 was detected in order to study its mechanism.Results:BMP9 was not only increased the activity of ALP on days 5,7,the calcium precipitation on 14 day was also increased compared with Ad-GFP group.miR-30a reduced the expression of ALP on BMP9-induced osteogenic differentiation.The expression of OPN and calcium precipitation were also inhibited by miR-30a.Conclusion:BMP9-induced osteogenic differentiation is partially inhibited by miR-30a in mesenchymal stem cell line C3H10 T1 /2.