MiR-204-5p/FOXC1/GDF7 Axis Regulates Osteogenic Differentiation of Human Adipose-Derived Stem Cells Via the AKT and p38 Signaling Pathways
Abstract Background: Human adipose-derived stem cells (hADSCs) are stem cells with the potential to differentiate in multiple directions. MiR-204-5p is poorly expressed during osteogenic differentiation of hADSCs, and its specific regulatory mechanism remains unclear. Here, we aimed to explore the function and possible molecular mechanism of miR-204-5p involved in the osteogenic differentiation of hADSCs. Methods: The expression pattern of miR-204-5p, Runx2, Alkaline phosphatase (ALP), Osteocalcin (OCN), and Forkhead box C1(FOXC1) and growth differentiation factor 7(GDF7) in hADSCs during osteogenesis were detected by qRT-PCR. Then, ALP and alizarin red staining (ARS) were used to detect the activity of osteoblasts and mineral deposition. Western blot was conducted to confirm the protein levels. The regulation relationship among miR-204-5p, FOXC1 and GDF7 was verified by double luciferase activity and CHIP experiment.Results: First, miR-204-5p expression was down-regulated and overexpressed miR-204-5p suppressed the osteogenic differentiation. Furthermore, the levels of FOXC1 and GDF7 were decreased in the miR-204-5p mimics group, which indicate that overexpressed miR-204-5p would suppress the expression of FOXC1 and GDF7 through binding the 3’UTR region each. Overexpression of FOXC1 or GDF7 could improve the inhibition of osteogenic differentiation of hADSCs induced by the miR-204-5p mimics. Moreover, FOXC1 could bind to the promoter of miR-204-5p and GDF7, promote the deacetylation of miR-204-5p and reduced the expression of miR-204-5p, thus promoting the expression of GDF7 during osteogenic differentiation. GDF7 could induce hADSCs osteogenesis differentiation by activating the AKT and P38 signaling pathways. Conclusions: Our results demonstrated that miR-204-5p/FOXC1/GDF7 axis regulates osteogenic differentiation of hADSCs via the AKT and p38 signaling pathways. This study further understood the regulatory mechanism of hADSCs differentiation balance from the perspective of miRNAs regulation.