Chidamide Promotes Osteogenic Differentiation of Bone Marrow Mesenchymal Stromal Cells from Patients with Myelodysplastic Syndromes
Objective:To explore the effects and mechanisms of chidamide on the osteogenic differentiation of bone marrow mesenchymal stromal cells(MSC)from myelodysplastic syndromes(MDS).Methods:MSC were isolated and cultured from bone marrow of MDS patients and healthy donors.CCK-8 assay was used to detect the effects of chidamide on the proliferation of MSC.The effects of chidamide on the activity of histone deacetylase(HD AC)in MSC was measured by a fluorescence assay kit and Western blot.Alkaline phosphatase(ALP)activity was detected on day 3 and calcium nodule formation was observed by Alizarin Red staining on day 21 after osteogenic differentiation.The expression of early and late osteogenic genes was detected on day 7 and day 21,respectively.RT-PCR and Western blot were used to detect the effects of chidamide on mRNA and protein expression of RUNX2 which is the key transcription factor during osteogenesis.Results:As the concentration of chidamide increased,the proliferation of MSC was inhibited.However,at a low concentration(1 μmol/L),chidamide had no significant inhibitory effect on MSC proliferation but significantly inhibited HD AC activity.In MSC from both MDS patients and healthy donors,chidamide(1 μmol/L)significantly increased ALP activity,calcium nodule formation,thereby mRNA expression of osteogenic genes,and restored the reduced osteogenic differentiation ability of MDS-MSC compared to normal MSC.Mechanistic studies showed that the osteogenic-promoting effect of chidamide may be related to the upregulation of RUNX2.Conclusion:Chidamide can inhibit HD AC activity in MSC,upregulate the expression of the osteogenic transcription factor RUNX2,and promote the osteogenic differentiation of MDS-MSC.
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