miR-615-3p inhibits osteogenic differentiation of adipose derived stem cells by negatively regulating the expression of FBLN1
Objective:To explore the function and mutual regulatory mechanism of miR-615-3p and its target gene fibulin 1(FBLN1)in osteogenic differentiation of adipose derived stem cells(ADSCs).Methods:Using lentivirus transfection to obtain miR-615-3p knockdown ADSCs and FBLN1 overexpressing ADSCs,Western blot and qRT-PCR were used to detect the knockdown efficien-cy of miR-615-3p and FBLN1 overexpression efficiency,as well as the expression of FBLN1 after knocking down miR-615-3p.The effects of knocking down miR-615-3p and overexpressing FBLN1 on the osteogenic differentiation ability of ADSCs were evaluated by al-kaline phosphatase(ALP)activity assay and alizarin red staining.Detection of gene expression of osteogenic cell specific transcription factors Osterix(OSX),dentin sialophosphoprotein(DSPP),and dentin matrix protein-1(DMP1)in knocking down miR-615-3p and overexpressing FBLN1 ADSCs by qRT-PCR.The expression changes of DMP1 and DSPP were detected by Western blot in knocking down miR-615-3p and overexpression of FBLN1.Results:After knocking down miR-615-3p in ADSCs,miR-615-3p gene expression decreased,ALP activity increased(P＜0.05),mineralized nodules increased,OSX,DMP1,DSPP gene expression levels increased(P＜0.05),and DMP1 and DSPP protein expression increased.The expression of FBLN1 was increased in ADSCs with low expression of miR-615-3p.After overexpression of FBLN1,the expression of FBLN1 gene increased,ALP activity increased(P＜0.05),mineral-ized nodules increased,and the expression levels of osteogenic related indicators such as OSX,DMP1,DSPP increased(P＜0.05).Conclusions:miR-615-3p inhibits osteogenic differentiation of ADSCs by downregulating FBLN1.
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